In view of the problems of the traditional cell incubator,such as the small range of cell culture types,the inability to adjust the internal space of the incubator according to needs,and the inconvenient sampling,this...In view of the problems of the traditional cell incubator,such as the small range of cell culture types,the inability to adjust the internal space of the incubator according to needs,and the inconvenient sampling,this study innovatively designed a cell incubator structure.It proposed a new design concept that can solve the above-mentioned shortcomings.The cell incubator after the new structural modification can adjust the internal space structure of cell culture by setting the bolt-fixed connection between the fixed plate and the vessel divider.It realizes the cultivation of various cells through refrigeration modules and heating modules.Through setting a sampling hole in the glass inner door,it is favorable for operators to take samples,making cell culture more convenient and efficient.展开更多
In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were in...In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were induced to form callus when cultured on vermicompost extract media along with coelomic fluid. Suspension medium was developed using vermicompost extract and coelomic fluid in 3:1 ratio. Phytochemical analysis of the alkaloid berberine was confirmed from callus, suspension cell culture and suspension medium by Thin Layer Chromatography and High Performance Liquid Chromatography. Vermicompost and its extracts with coelomic fluid have shown maximum (100 per cent) response of callus induction. Callus mass enlarged with increasing concentration of coelomic fluid and callus growth was assessed from the biomass. Incubation of culture tubes in dark supported callus development significantly. The Rf value of 0.36 confirmed the presence of berberine by Thin Layer Chromatography. Qualitative analysis confirmed the presence of alkaloid berberine with the retention time of 2.8 minutes similar to that of standard reference sample from Sigma chemicals, USA. The suspension medium turned deep yellow because of the release of the alkaloid. Vermicompost and its extracts along with coelomic fluid have shown the economical approach for micropropagation of economically and medicinally important plants.展开更多
Plant cell culture technology is a technology that applies the research results of cell engineering to produce plant biological products at the cellular level.In recent years,the secondary metabolites of plants have a...Plant cell culture technology is a technology that applies the research results of cell engineering to produce plant biological products at the cellular level.In recent years,the secondary metabolites of plants have attracted more and more attention.The use of plant cell culture technology is a fast and efficient method of producing secondary metabolites.展开更多
Three-dimensional(3D)culture systems are becoming increasingly popular due to their ability to mimic tissue-like structures more effectively than the monolayer cultures.In cancer and stem cell research,the natural cel...Three-dimensional(3D)culture systems are becoming increasingly popular due to their ability to mimic tissue-like structures more effectively than the monolayer cultures.In cancer and stem cell research,the natural cell characteristics and architectures are closely mimicked by the 3D cell models.Thus,the 3D cell cultures are promising and suitable systems for various proposes,ranging from disease modeling to drug target identification as well as potential therapeutic substances that may transform our lives.This review provides a comprehensive compendium of recent advancements in culturing cells,in particular cancer and stem cells,using 3D culture techniques.The major approaches highlighted here include cell spheroids,hydrogel embedding,bioreactors,scaffolds,and bioprinting.In addition,the progress of employing 3D cell culture systems as a platform for cancer and stem cell research was addressed,and the prominent studies of 3D cell culture systems were discussed.展开更多
Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discov...Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discovery process is still challenging due to difficulties in controlling the stem cell microenvironment and the limited availability of high-throughput systems.Recently,researchers have been actively developing and evaluating three-dimensional(3D)cell culture-based platforms using microfluidic technologies,such as organ-on-a-chip and organoid-on-a-chip platforms,and they have achieved promising breakthroughs in stem cell engineering.In this review,we start with a comprehensive discussion on the importance of microfluidic 3D cell culture techniques in stem cell research and their technical strategies in the field of drug discovery.In a subsequent section,we discuss microfluidic 3D cell culture techniques for high-throughput analysis for use in stem cell research.In addition,some potential and practical applications of organ-on-a-chip or organoid-on-a-chip platforms using stem cells as drug screening and disease models are highlighted.展开更多
A comparative study was carried out with ethanolic (80%) extracts from leaves and cell cultures of three Callistemon species, namely C. lanceolatus (CL), C. viridiflorous (CV), and C. comboynensis (CC). Cell suspensio...A comparative study was carried out with ethanolic (80%) extracts from leaves and cell cultures of three Callistemon species, namely C. lanceolatus (CL), C. viridiflorous (CV), and C. comboynensis (CC). Cell suspensions of the three species were grown in liquid Murashige and Skoog (MS) medium (100 ml) supplemented with 0.9 mg·g-1 kinetin in combination with 1.1 mg·g-1 NAA. The CL leaf extract was standardized to contain the highest amount of phenolics (104 ± 2.0 mg·g-1), followed by CC (95.8 ± 1.2 mg·g-1) and CV (79.8 ± 4.6 mg·g-1). On the other hand, cell cultures of CV contained more phenolics (14.9 ± 0.6 mg·g-1) than those of the other two species, CL and CC, which contained 12.2 ± 0.16 and 9.12 ± 0.16 mg·g-1, respectively. Nevertheless, CV leaf extract exhibited the highest antioxidant activity (91.4% ± 0.4%) at a concentration of 1000 μg·ml-1, comparable to 100 μg·ml-1 gallic acid (90.8% ± 1.5%).展开更多
Studying physiological and pathophysiological mechanisms in the liver on a molecular basis is a challenging task.During two dimensional(2D) culture conditions hepatocytes dedifferentiate rapidly by losing metabolic fu...Studying physiological and pathophysiological mechanisms in the liver on a molecular basis is a challenging task.During two dimensional(2D) culture conditions hepatocytes dedifferentiate rapidly by losing metabolic functions and structural integrity.Hence,inappropriate 2D hepatocellular models hamper studies on the xenobiotic metabolism of the liver which strongly influences drug potency.Also,the lack of effective therapies against hepatocellular carcinoma shows the urgent need for robust models to investigate liver functions in a defined hepatic microenvironment.Here,we summarize and discuss three-dimensional cultures of hepatocytes,herein referred to as hepatospheres,which provide versatile tools to investigate hepatic metabolism,stemness and cancer development.展开更多
AIM:We compared polymerase chain reaction(PCR)to cell culture isolation for the laboratory diagnosis of ocular herpes simplex virus(HSV)disease. ? METHODS:Laboratory and medical records of consecutive patients were re...AIM:We compared polymerase chain reaction(PCR)to cell culture isolation for the laboratory diagnosis of ocular herpes simplex virus(HSV)disease. ? METHODS:Laboratory and medical records of consecutive patients were reviewed for results of 1)HSV PCR testing,2) HSV cell culture isolation,and 3)clinical diagnosis.PCR results were statistically compared to cell culture isolation and patients initially diagnosed for ocular HSV infection. RESULTS:Of 581 cases submitted for laboratory testing, 520 were PCR negative,cell culture negative(89.6%);0 were PCR negative,cell culture positive(0%);27 were PCR positive,cell culture negative(4.6%);and 34 were PCR positive,cell culture positive(5.8%).PCR tested more positive than cell culture isolation(McNemar’s,P=0.0001).Of 47 HSV PCR positive cases with complete medical records,19 were cell culture negative for HSV and 28 were cell culture positive for HSV.Fourteen of 19 cell culture negative cases (74%)(Without PCR,5 cases of HSV would be missed)and 25 of the 28 cell culture positive cases(89%)(Laboratory testing was necessary for diagnosing 3 cases)were clinically diagnosed with HSV at the initial examination. CONCLUSION:PCR was a more definitive test for diagnosing HSV ocular infection than cell culture isolation.Cell culture isolation alone can miss an atypical presentation of HSV ocular infection.展开更多
Human pluripotent stem cells(hPSCs)are important resources for cell-based therapies and pharmaceutical applications.In order to realize the potential of hPSCs,it is critical to develop suitable technologies required f...Human pluripotent stem cells(hPSCs)are important resources for cell-based therapies and pharmaceutical applications.In order to realize the potential of hPSCs,it is critical to develop suitable technologies required for specific applications.Most hPSC technologies depend on cell culture,and are critically influenced by culture medium composition,extracellular matrices,handling methods,and culture platforms.This review summarizes the major technological advances in hPSC culture,and highlights the opportunities and challenges in future therapeutic applications.展开更多
The development and delivery of high quality therapeutic products necessitates the need for highthrough-put (HTP) process development tools. Traditionally, these works requires a combination of shake flask and small-s...The development and delivery of high quality therapeutic products necessitates the need for highthrough-put (HTP) process development tools. Traditionally, these works requires a combination of shake flask and small-scale stirred tank bioreactor (STR) which are labor and resource intensive and time-consuming. Here we demonstrate a strategy for rapid and robust cell culture process development by evaluating and implementing the use of a new HTP disposable micro bioreactor (MBR) called AMBRTM system (Advanced Microscale Bioreactor) that has the capabilities for automated sampling, feed addition, pH, dissolved oxygen (DO), gassing and agitation controls. In these studies the performance of two monoclonal antibody (MAb) producing cell lines (MAb1 and MAb2) was evaluated both in the AMBR system and 3-L STR. We demonstrated that cell culture performance (growth and viability, production titer and product quality) were similar in both vessel systems. Furthermore, process control and feed optimization were demonstrated in an additional cell line (MAb3) in the disposable MBR and its performance confirmed at STR scale. The results indicate that the AMBR system can be used to streamline the process development effort and facilitate a rapid and robust cell culture process development effort for MAb programs in a HTP manner.展开更多
Metastatic Merkel Cell carcinoma (MCC) is a highly unusual and aggressive skin cancer that presents as a small, pink to violet skin lesion and metastasizes early in its growth. Metastatic MCC is generally treated with...Metastatic Merkel Cell carcinoma (MCC) is a highly unusual and aggressive skin cancer that presents as a small, pink to violet skin lesion and metastasizes early in its growth. Metastatic MCC is generally treated with small cell lung cancer chemotherapy regimens, because the tumor consists of neuroendocrine cells, but patients generally do not have durable responses. The pathogenesis of MCC has recently been attributed to the Merkel Cell polyoma virus. This virus activates the cellular retinoblastoma oncoprotein and cell cycle machinery, triggering continual cellular proliferation. A 77-year-old man developed extensive MCC metastases, involving more than one fourth of his scalp and numerous cervical lymph nodes. Following failure of initial chemotherapy and radiation, effective palliation was achieved by using a sequence of electron-beam radiotherapy, low dose gemcitabine, and etoposide, resulting in significant periods of tumor regression and prolonged survival. A novel circulating tumor cell (CTC) culture assay was performed on four separate clinic visits during the treatment period. Tumor colonies were cultured from the patient’s peripheral blood and CTC colony counts were correlated with clinical treatment response. Not only did the patient respond to palliative cell cycle directed chemotherapy and electron beam radiation, but we demonstrated that CTC can be cultured from peripheral blood of MCC patients and serve as a predictive marker to monitor treatment response.展开更多
Background: Increased mitogen-activated protein kinase (MAPK) phosphorylation has been detected in peripheral nerve of human subjects and animal models with diabetes as well as high-glucose exposed human Schwann cells...Background: Increased mitogen-activated protein kinase (MAPK) phosphorylation has been detected in peripheral nerve of human subjects and animal models with diabetes as well as high-glucose exposed human Schwann cells, and have been implicated in diabetic peripheral neuropathy. In our recent studies, leukocytetype 12/15-lipoxygenase inhibition or gene deficiency alleviated large and small nerve fiber dysfunction, but not intraepidermal nerve fiber loss in streptozotocin-diabetic mice. Methods: To address a mechanism we evaluated the potential for pharmacological 12/15-lipoxygenase inhibition to counteract excessive MAPK phosphorylation in mouse and cell culture models of diabetic neuropathy. C57Bl6/J mice were made diabetic with streptozotocin and maintained with or without the 12/15-lipoxygenase inhibitor cinnamyl-3,4-dihydroxy-α-cyanocinnamate (CDC). Human Schwann cells were cultured in5.5 mMor30 mMglucose with or without CDC. Results: 12(S) HETE concentrations (ELISA), as well as 12/15-lipoxygenase expression and p38 MAPK, ERK, and SAPK/JNK phosphorylation (all by Western blot analysis) were increased in the peripheral nerve and spinal cord of diabetic mice as well as in high glucose-exposed human Schwann cells. CDC counteracted diabetes-induced increase in 12(S)HETE concentrations (a measure of 12/15-lipoxygenase activity), but not 12/15-lipoxygenase overexpression, in sciatic nerve and spinal cord. The inhibitor blunted excessive p38 MAPK and ERK, but not SAPK/ JNK, phosphorylation in sciatic nerve and high glucose exposed human Schwann cells, but did not affect MAPK, ERK, and SAPK/JNK phosphorylation in spinal cord. Conclusion: 12/15-lipoxygenase inhibition counteracts diabetes related MAPK phosphorylation in mouse and cell culture models of diabetic neuropathy and implies that 12/15-lipoxygenase inhibitors may be an effective treatment for diabetic peripheral neuropathy.展开更多
Multicellular layers(MCLs) have previously been used to determine the pharmacokinetics of a variety of different cancer drugs including paclitaxel, doxorubicin, methotrexate, and 5-fluorouracil across a number of cell...Multicellular layers(MCLs) have previously been used to determine the pharmacokinetics of a variety of different cancer drugs including paclitaxel, doxorubicin, methotrexate, and 5-fluorouracil across a number of cell lines. It is not known how nanoparticles(NPs) navigate through the tumor microenvironment once they leave the tumor blood vessel.In this study, we used the MCL model to study the uptake and penetration dynamics of NPs. Gold nanoparticles(GNPs)were used as a model system to map the NP distribution within tissue-like structures. Our results show that NP uptake and transport are dependent on the tumor cell type. MDA-MB-231 tissue showed deeper penetration of GNPs as compared to MCF-7 one. Intracellular and extracellular distributions of NPs were mapped using Cyto Viva imaging. The ability of MCLs to mimic tumor tissue characteristics makes them a useful tool in assessing the efficacy of particle distribution in solid tumors.展开更多
Stem cells (SCs), the undifferentiated biological cells, have the infinite capacity to self-renew and the pluripotent ability to differentiate. SCs and their derived products offer great promise for biomedical applica...Stem cells (SCs), the undifferentiated biological cells, have the infinite capacity to self-renew and the pluripotent ability to differentiate. SCs and their derived products offer great promise for biomedical applications such as cell therapy, tissue engineering, regenerative medicine and drug screening. However, the clinical applications of SCs require a large amount of SCs with high quality and the number of SCs from their tissue resources is very limited. Large-scale expansion is needed to generate homogeneous SCs with good biological characteristics for clinical application. This necessitates a bioreactor system to provide controllable and stable conditions for stem cell (SC) culture. Traditional methods of bioreactor for maintenance and expansion of cells rely on two-dimensional (2-D) culture techniques, leading to loss self-renewal ability and differentiation capacity upon long-term culture. New approaches for SC expansion with bioreactor employ three-dimensional (3-D) cell growth to mimic their environment in vivo. In this review, we summarize the application of bioreactors in SC culture.展开更多
In model experiments were studied the effect of cosmo-geophysical factors of environment (hypomagnetic conditions during 2 days ≈ 1 mkT;electromagnetic irradiation (10 min - 2 MHz with amplitude 5 V/m and power 30 mk...In model experiments were studied the effect of cosmo-geophysical factors of environment (hypomagnetic conditions during 2 days ≈ 1 mkT;electromagnetic irradiation (10 min - 2 MHz with amplitude 5 V/m and power 30 mkVt, background 2 - 4 mkVt), γ-quantum (10 min—from the source 137Cs) and its combined effect on the physic-chemical properties (ORP and pH) of growing medium for cell culture of mammals as nutrition medium 199 (PanEco, Russia). It was used a clear solution of medium (solution 1) and with the adding of 10% embryo bull serum—model of bio-medium (solution 2). Hypomagnetic conditions evoked the decreasing of ORP and pH value in both solutions, electromagnetic irradiation in the solution 1 which evoked the decreasing of ORP and the increasing of pH value, and in the solution 2, on the contrary, the increasing of ORP with the unchanging pH value. γ-radiation sharply decreased ORP value and didn’t change pH in solution 1, i.e. the reduction properties increased. There is insignificant increasing of ORP value and the decreasing of pH is noted in the solution 2, that it is characterized with the increasing of oxidative properties of solution. Under the combined effect of hypomagnetic conditions and electromagnetic irradiation, the values of investigating parameters in the solution 1 decreased and in the solution 2 increased. It was observed acute decreasing of ORP value in both solutions under the combined effect of hypomagnetic conditions and γ-radiation, i.e. the reductive properties of the solutions increased sharply. In this the concentration H+ significantly decreased, (p γ-radiation led to the decreasing of ORP and pH values in both solutions. Thus, the studying factors significantly change the oxidation-reduction properties of growing mediums. The investigation of the processes in biological mediums plays the important role in the assessment of environment effect during the flight in inter-planet space.展开更多
At the present time animal cell culture is more significant and multifarious application tool for current research streams. A lot of field assorted from animal cell culture such: stem cell biology, IVF technology, can...At the present time animal cell culture is more significant and multifarious application tool for current research streams. A lot of field assorted from animal cell culture such: stem cell biology, IVF technology, cancer cell biology, monoclonal antibody production, recombinant protein production, gene therapy, vaccine manufacturing, novel drug selection and improvement. In this review conclude animal cell culture as well as its展开更多
Some biological characteristics of human bone marrow mesenchymal stem cells (MSCs) cultured in vitro were observed. hMSCs were isolated from bone marrow and purified by density gradient centrifugation method, and then...Some biological characteristics of human bone marrow mesenchymal stem cells (MSCs) cultured in vitro were observed. hMSCs were isolated from bone marrow and purified by density gradient centrifugation method, and then cultured in vitro. The proliferation and growth characteristics of hMSCs were observed in primary and passage culture. MSCs of passage 3 were examined for the purify by positive rate of CD29 and CD44 through flow cytometry. Human bone marrow MSCs showed active proliferation capacity in vitro. The purify of MSCs separated by our method was higher than 90 %. It was concluded that hMSCs have been successfully cultured and expanded effectively. It provided a foundation for further investigation and application of MSCs.展开更多
This short review is to list pros and cons which are based on the literature and personal experience in cell culture studies related to possible commercial production of artificial meat as functional food. The general...This short review is to list pros and cons which are based on the literature and personal experience in cell culture studies related to possible commercial production of artificial meat as functional food. The general view of muscle composition and determinants of meat quality are shortly described. Principles of muscle cell propagation in culture and mutual relationships between different cell types present in this organ are briefly discussed. Additionally, the effects of some cytokines and growth factors for muscle cell growth and muscle tissue development are indicated. Finally, conclusion remarks related to detrimental consequences of meat production to natural environment as well as personal opinion of author on the prospects of artificial meat production are declared.展开更多
This study aimed to establish a new in vitro three-dimensional (3D) cell culture and use quantum dots (QDs) molecular imaging to examine the invasive behaviors of hepatocellular carcinoma (HCC) cells.Each well of the ...This study aimed to establish a new in vitro three-dimensional (3D) cell culture and use quantum dots (QDs) molecular imaging to examine the invasive behaviors of hepatocellular carcinoma (HCC) cells.Each well of the 24-well cell culture plate was cover-slipped.Matrigel diluted with serum-free DMEM was added and HCCLM9 cells were cultured on the Matrigel.The cell morphological and cell growth characteristics were observed by inverted microscopy and laser confocal microscopy at different culture time.Cell invasive features were monitored by QDs-based real-time molecular imaging techniques.The results showed that on this 3D cell culture platform,HCCLM9 cells exhibited typical multi-step invasive behaviors,including reversion of cell senescence,active focal proliferation and dominant clones invasion.During the process,cells under 3D cell culture showed biological behaviors of spatio-temporal characteristics.Cells first merged on the surface of matrix,then gradually infiltrated and migrated into deep part of matrix,presenting polygonal morphology with stretched protrusions,forming tubular,annular and even network structure,which suggested that HCC cells have the morphological basis for vasculogenic mimicry.In addition,small cell clones with their edges well-circumscribed in early stage,progressed into a large irregular clone with ill-defined edge,while the other cells developed invadopodia.And QDs probing showed MT1-MMP was strongly expressed in the invadopodia.These findings indicate that a novel 3D cell culture platform has been successfully established,which can mimic the in vivo tumor microenvironment,and when combined with QDs-based molecular imaging,it can help to better investigate the invasive behaviors of HCC cells.展开更多
基金Supported by Young and Middle-aged Teacher Education Research Project of Fujian Province(Science and Technology Category:JAT210477)College Student Innovation and Entrepreneurship Training Program of Xiamen Medical College(X202112631068)。
文摘In view of the problems of the traditional cell incubator,such as the small range of cell culture types,the inability to adjust the internal space of the incubator according to needs,and the inconvenient sampling,this study innovatively designed a cell incubator structure.It proposed a new design concept that can solve the above-mentioned shortcomings.The cell incubator after the new structural modification can adjust the internal space structure of cell culture by setting the bolt-fixed connection between the fixed plate and the vessel divider.It realizes the cultivation of various cells through refrigeration modules and heating modules.Through setting a sampling hole in the glass inner door,it is favorable for operators to take samples,making cell culture more convenient and efficient.
文摘In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were induced to form callus when cultured on vermicompost extract media along with coelomic fluid. Suspension medium was developed using vermicompost extract and coelomic fluid in 3:1 ratio. Phytochemical analysis of the alkaloid berberine was confirmed from callus, suspension cell culture and suspension medium by Thin Layer Chromatography and High Performance Liquid Chromatography. Vermicompost and its extracts with coelomic fluid have shown maximum (100 per cent) response of callus induction. Callus mass enlarged with increasing concentration of coelomic fluid and callus growth was assessed from the biomass. Incubation of culture tubes in dark supported callus development significantly. The Rf value of 0.36 confirmed the presence of berberine by Thin Layer Chromatography. Qualitative analysis confirmed the presence of alkaloid berberine with the retention time of 2.8 minutes similar to that of standard reference sample from Sigma chemicals, USA. The suspension medium turned deep yellow because of the release of the alkaloid. Vermicompost and its extracts along with coelomic fluid have shown the economical approach for micropropagation of economically and medicinally important plants.
文摘Plant cell culture technology is a technology that applies the research results of cell engineering to produce plant biological products at the cellular level.In recent years,the secondary metabolites of plants have attracted more and more attention.The use of plant cell culture technology is a fast and efficient method of producing secondary metabolites.
文摘Three-dimensional(3D)culture systems are becoming increasingly popular due to their ability to mimic tissue-like structures more effectively than the monolayer cultures.In cancer and stem cell research,the natural cell characteristics and architectures are closely mimicked by the 3D cell models.Thus,the 3D cell cultures are promising and suitable systems for various proposes,ranging from disease modeling to drug target identification as well as potential therapeutic substances that may transform our lives.This review provides a comprehensive compendium of recent advancements in culturing cells,in particular cancer and stem cells,using 3D culture techniques.The major approaches highlighted here include cell spheroids,hydrogel embedding,bioreactors,scaffolds,and bioprinting.In addition,the progress of employing 3D cell culture systems as a platform for cancer and stem cell research was addressed,and the prominent studies of 3D cell culture systems were discussed.
基金supported by the National Research Foundation of Korea (NRF) (NRF2017R1C1B2002377, NRF-2016R1A5A1010148, and NRF2019R1A2C1003111)funded by the Ministry of Science and ICT (MSIT)partly supported by the Technology Innovation Program (No.10067787)funded by the Ministry of Trade, Industry & Energy (MOTE, Korea)
文摘Although the recent advances in stem cell engineering have gained a great deal of attention due to their high potential in clinical research,the applicability of stem cells for preclinical screening in the drug discovery process is still challenging due to difficulties in controlling the stem cell microenvironment and the limited availability of high-throughput systems.Recently,researchers have been actively developing and evaluating three-dimensional(3D)cell culture-based platforms using microfluidic technologies,such as organ-on-a-chip and organoid-on-a-chip platforms,and they have achieved promising breakthroughs in stem cell engineering.In this review,we start with a comprehensive discussion on the importance of microfluidic 3D cell culture techniques in stem cell research and their technical strategies in the field of drug discovery.In a subsequent section,we discuss microfluidic 3D cell culture techniques for high-throughput analysis for use in stem cell research.In addition,some potential and practical applications of organ-on-a-chip or organoid-on-a-chip platforms using stem cells as drug screening and disease models are highlighted.
文摘A comparative study was carried out with ethanolic (80%) extracts from leaves and cell cultures of three Callistemon species, namely C. lanceolatus (CL), C. viridiflorous (CV), and C. comboynensis (CC). Cell suspensions of the three species were grown in liquid Murashige and Skoog (MS) medium (100 ml) supplemented with 0.9 mg·g-1 kinetin in combination with 1.1 mg·g-1 NAA. The CL leaf extract was standardized to contain the highest amount of phenolics (104 ± 2.0 mg·g-1), followed by CC (95.8 ± 1.2 mg·g-1) and CV (79.8 ± 4.6 mg·g-1). On the other hand, cell cultures of CV contained more phenolics (14.9 ± 0.6 mg·g-1) than those of the other two species, CL and CC, which contained 12.2 ± 0.16 and 9.12 ± 0.16 mg·g-1, respectively. Nevertheless, CV leaf extract exhibited the highest antioxidant activity (91.4% ± 0.4%) at a concentration of 1000 μg·ml-1, comparable to 100 μg·ml-1 gallic acid (90.8% ± 1.5%).
基金Supported by the Austrian Science Fund,FWF,NO.P19598-B13 and SFB F28,the"Hochschuljubilumsstiftung der Stadt Wien",the Herzfelder Family Foundationthe European Union,FP7 Health Research,NO.HEALTH-F4-2008-202047
文摘Studying physiological and pathophysiological mechanisms in the liver on a molecular basis is a challenging task.During two dimensional(2D) culture conditions hepatocytes dedifferentiate rapidly by losing metabolic functions and structural integrity.Hence,inappropriate 2D hepatocellular models hamper studies on the xenobiotic metabolism of the liver which strongly influences drug potency.Also,the lack of effective therapies against hepatocellular carcinoma shows the urgent need for robust models to investigate liver functions in a defined hepatic microenvironment.Here,we summarize and discuss three-dimensional cultures of hepatocytes,herein referred to as hepatospheres,which provide versatile tools to investigate hepatic metabolism,stemness and cancer development.
文摘AIM:We compared polymerase chain reaction(PCR)to cell culture isolation for the laboratory diagnosis of ocular herpes simplex virus(HSV)disease. ? METHODS:Laboratory and medical records of consecutive patients were reviewed for results of 1)HSV PCR testing,2) HSV cell culture isolation,and 3)clinical diagnosis.PCR results were statistically compared to cell culture isolation and patients initially diagnosed for ocular HSV infection. RESULTS:Of 581 cases submitted for laboratory testing, 520 were PCR negative,cell culture negative(89.6%);0 were PCR negative,cell culture positive(0%);27 were PCR positive,cell culture negative(4.6%);and 34 were PCR positive,cell culture positive(5.8%).PCR tested more positive than cell culture isolation(McNemar’s,P=0.0001).Of 47 HSV PCR positive cases with complete medical records,19 were cell culture negative for HSV and 28 were cell culture positive for HSV.Fourteen of 19 cell culture negative cases (74%)(Without PCR,5 cases of HSV would be missed)and 25 of the 28 cell culture positive cases(89%)(Laboratory testing was necessary for diagnosing 3 cases)were clinically diagnosed with HSV at the initial examination. CONCLUSION:PCR was a more definitive test for diagnosing HSV ocular infection than cell culture isolation.Cell culture isolation alone can miss an atypical presentation of HSV ocular infection.
基金Supported by University of Macao Multi-Year Research Grants,No.MYRG2015-00228-FHS and MYRG2018-00135-FHSMacao Science and Technology Development Fund,No.FDCT/131/2014/A3 and FDCT/056/2015/A2
文摘Human pluripotent stem cells(hPSCs)are important resources for cell-based therapies and pharmaceutical applications.In order to realize the potential of hPSCs,it is critical to develop suitable technologies required for specific applications.Most hPSC technologies depend on cell culture,and are critically influenced by culture medium composition,extracellular matrices,handling methods,and culture platforms.This review summarizes the major technological advances in hPSC culture,and highlights the opportunities and challenges in future therapeutic applications.
文摘The development and delivery of high quality therapeutic products necessitates the need for highthrough-put (HTP) process development tools. Traditionally, these works requires a combination of shake flask and small-scale stirred tank bioreactor (STR) which are labor and resource intensive and time-consuming. Here we demonstrate a strategy for rapid and robust cell culture process development by evaluating and implementing the use of a new HTP disposable micro bioreactor (MBR) called AMBRTM system (Advanced Microscale Bioreactor) that has the capabilities for automated sampling, feed addition, pH, dissolved oxygen (DO), gassing and agitation controls. In these studies the performance of two monoclonal antibody (MAb) producing cell lines (MAb1 and MAb2) was evaluated both in the AMBR system and 3-L STR. We demonstrated that cell culture performance (growth and viability, production titer and product quality) were similar in both vessel systems. Furthermore, process control and feed optimization were demonstrated in an additional cell line (MAb3) in the disposable MBR and its performance confirmed at STR scale. The results indicate that the AMBR system can be used to streamline the process development effort and facilitate a rapid and robust cell culture process development effort for MAb programs in a HTP manner.
文摘Metastatic Merkel Cell carcinoma (MCC) is a highly unusual and aggressive skin cancer that presents as a small, pink to violet skin lesion and metastasizes early in its growth. Metastatic MCC is generally treated with small cell lung cancer chemotherapy regimens, because the tumor consists of neuroendocrine cells, but patients generally do not have durable responses. The pathogenesis of MCC has recently been attributed to the Merkel Cell polyoma virus. This virus activates the cellular retinoblastoma oncoprotein and cell cycle machinery, triggering continual cellular proliferation. A 77-year-old man developed extensive MCC metastases, involving more than one fourth of his scalp and numerous cervical lymph nodes. Following failure of initial chemotherapy and radiation, effective palliation was achieved by using a sequence of electron-beam radiotherapy, low dose gemcitabine, and etoposide, resulting in significant periods of tumor regression and prolonged survival. A novel circulating tumor cell (CTC) culture assay was performed on four separate clinic visits during the treatment period. Tumor colonies were cultured from the patient’s peripheral blood and CTC colony counts were correlated with clinical treatment response. Not only did the patient respond to palliative cell cycle directed chemotherapy and electron beam radiation, but we demonstrated that CTC can be cultured from peripheral blood of MCC patients and serve as a predictive marker to monitor treatment response.
文摘Background: Increased mitogen-activated protein kinase (MAPK) phosphorylation has been detected in peripheral nerve of human subjects and animal models with diabetes as well as high-glucose exposed human Schwann cells, and have been implicated in diabetic peripheral neuropathy. In our recent studies, leukocytetype 12/15-lipoxygenase inhibition or gene deficiency alleviated large and small nerve fiber dysfunction, but not intraepidermal nerve fiber loss in streptozotocin-diabetic mice. Methods: To address a mechanism we evaluated the potential for pharmacological 12/15-lipoxygenase inhibition to counteract excessive MAPK phosphorylation in mouse and cell culture models of diabetic neuropathy. C57Bl6/J mice were made diabetic with streptozotocin and maintained with or without the 12/15-lipoxygenase inhibitor cinnamyl-3,4-dihydroxy-α-cyanocinnamate (CDC). Human Schwann cells were cultured in5.5 mMor30 mMglucose with or without CDC. Results: 12(S) HETE concentrations (ELISA), as well as 12/15-lipoxygenase expression and p38 MAPK, ERK, and SAPK/JNK phosphorylation (all by Western blot analysis) were increased in the peripheral nerve and spinal cord of diabetic mice as well as in high glucose-exposed human Schwann cells. CDC counteracted diabetes-induced increase in 12(S)HETE concentrations (a measure of 12/15-lipoxygenase activity), but not 12/15-lipoxygenase overexpression, in sciatic nerve and spinal cord. The inhibitor blunted excessive p38 MAPK and ERK, but not SAPK/ JNK, phosphorylation in sciatic nerve and high glucose exposed human Schwann cells, but did not affect MAPK, ERK, and SAPK/JNK phosphorylation in spinal cord. Conclusion: 12/15-lipoxygenase inhibition counteracts diabetes related MAPK phosphorylation in mouse and cell culture models of diabetic neuropathy and implies that 12/15-lipoxygenase inhibitors may be an effective treatment for diabetic peripheral neuropathy.
文摘Multicellular layers(MCLs) have previously been used to determine the pharmacokinetics of a variety of different cancer drugs including paclitaxel, doxorubicin, methotrexate, and 5-fluorouracil across a number of cell lines. It is not known how nanoparticles(NPs) navigate through the tumor microenvironment once they leave the tumor blood vessel.In this study, we used the MCL model to study the uptake and penetration dynamics of NPs. Gold nanoparticles(GNPs)were used as a model system to map the NP distribution within tissue-like structures. Our results show that NP uptake and transport are dependent on the tumor cell type. MDA-MB-231 tissue showed deeper penetration of GNPs as compared to MCF-7 one. Intracellular and extracellular distributions of NPs were mapped using Cyto Viva imaging. The ability of MCLs to mimic tumor tissue characteristics makes them a useful tool in assessing the efficacy of particle distribution in solid tumors.
文摘Stem cells (SCs), the undifferentiated biological cells, have the infinite capacity to self-renew and the pluripotent ability to differentiate. SCs and their derived products offer great promise for biomedical applications such as cell therapy, tissue engineering, regenerative medicine and drug screening. However, the clinical applications of SCs require a large amount of SCs with high quality and the number of SCs from their tissue resources is very limited. Large-scale expansion is needed to generate homogeneous SCs with good biological characteristics for clinical application. This necessitates a bioreactor system to provide controllable and stable conditions for stem cell (SC) culture. Traditional methods of bioreactor for maintenance and expansion of cells rely on two-dimensional (2-D) culture techniques, leading to loss self-renewal ability and differentiation capacity upon long-term culture. New approaches for SC expansion with bioreactor employ three-dimensional (3-D) cell growth to mimic their environment in vivo. In this review, we summarize the application of bioreactors in SC culture.
文摘In model experiments were studied the effect of cosmo-geophysical factors of environment (hypomagnetic conditions during 2 days ≈ 1 mkT;electromagnetic irradiation (10 min - 2 MHz with amplitude 5 V/m and power 30 mkVt, background 2 - 4 mkVt), γ-quantum (10 min—from the source 137Cs) and its combined effect on the physic-chemical properties (ORP and pH) of growing medium for cell culture of mammals as nutrition medium 199 (PanEco, Russia). It was used a clear solution of medium (solution 1) and with the adding of 10% embryo bull serum—model of bio-medium (solution 2). Hypomagnetic conditions evoked the decreasing of ORP and pH value in both solutions, electromagnetic irradiation in the solution 1 which evoked the decreasing of ORP and the increasing of pH value, and in the solution 2, on the contrary, the increasing of ORP with the unchanging pH value. γ-radiation sharply decreased ORP value and didn’t change pH in solution 1, i.e. the reduction properties increased. There is insignificant increasing of ORP value and the decreasing of pH is noted in the solution 2, that it is characterized with the increasing of oxidative properties of solution. Under the combined effect of hypomagnetic conditions and electromagnetic irradiation, the values of investigating parameters in the solution 1 decreased and in the solution 2 increased. It was observed acute decreasing of ORP value in both solutions under the combined effect of hypomagnetic conditions and γ-radiation, i.e. the reductive properties of the solutions increased sharply. In this the concentration H+ significantly decreased, (p γ-radiation led to the decreasing of ORP and pH values in both solutions. Thus, the studying factors significantly change the oxidation-reduction properties of growing mediums. The investigation of the processes in biological mediums plays the important role in the assessment of environment effect during the flight in inter-planet space.
文摘At the present time animal cell culture is more significant and multifarious application tool for current research streams. A lot of field assorted from animal cell culture such: stem cell biology, IVF technology, cancer cell biology, monoclonal antibody production, recombinant protein production, gene therapy, vaccine manufacturing, novel drug selection and improvement. In this review conclude animal cell culture as well as its
文摘Some biological characteristics of human bone marrow mesenchymal stem cells (MSCs) cultured in vitro were observed. hMSCs were isolated from bone marrow and purified by density gradient centrifugation method, and then cultured in vitro. The proliferation and growth characteristics of hMSCs were observed in primary and passage culture. MSCs of passage 3 were examined for the purify by positive rate of CD29 and CD44 through flow cytometry. Human bone marrow MSCs showed active proliferation capacity in vitro. The purify of MSCs separated by our method was higher than 90 %. It was concluded that hMSCs have been successfully cultured and expanded effectively. It provided a foundation for further investigation and application of MSCs.
基金provided by grant from the National Science Centre in Poland (UMO-2013/11/B/NZ5/03106)
文摘This short review is to list pros and cons which are based on the literature and personal experience in cell culture studies related to possible commercial production of artificial meat as functional food. The general view of muscle composition and determinants of meat quality are shortly described. Principles of muscle cell propagation in culture and mutual relationships between different cell types present in this organ are briefly discussed. Additionally, the effects of some cytokines and growth factors for muscle cell growth and muscle tissue development are indicated. Finally, conclusion remarks related to detrimental consequences of meat production to natural environment as well as personal opinion of author on the prospects of artificial meat production are declared.
基金supported by grants from National Natural Science Foundation of China(No.81171396)Creative Research Groups of the National Natural Science Foundation of China(No.20921062)+1 种基金National Science and Technology Major Project(No.2012ZX10002012-12)National University Students Innovation Training Project of China(No.111048673)
文摘This study aimed to establish a new in vitro three-dimensional (3D) cell culture and use quantum dots (QDs) molecular imaging to examine the invasive behaviors of hepatocellular carcinoma (HCC) cells.Each well of the 24-well cell culture plate was cover-slipped.Matrigel diluted with serum-free DMEM was added and HCCLM9 cells were cultured on the Matrigel.The cell morphological and cell growth characteristics were observed by inverted microscopy and laser confocal microscopy at different culture time.Cell invasive features were monitored by QDs-based real-time molecular imaging techniques.The results showed that on this 3D cell culture platform,HCCLM9 cells exhibited typical multi-step invasive behaviors,including reversion of cell senescence,active focal proliferation and dominant clones invasion.During the process,cells under 3D cell culture showed biological behaviors of spatio-temporal characteristics.Cells first merged on the surface of matrix,then gradually infiltrated and migrated into deep part of matrix,presenting polygonal morphology with stretched protrusions,forming tubular,annular and even network structure,which suggested that HCC cells have the morphological basis for vasculogenic mimicry.In addition,small cell clones with their edges well-circumscribed in early stage,progressed into a large irregular clone with ill-defined edge,while the other cells developed invadopodia.And QDs probing showed MT1-MMP was strongly expressed in the invadopodia.These findings indicate that a novel 3D cell culture platform has been successfully established,which can mimic the in vivo tumor microenvironment,and when combined with QDs-based molecular imaging,it can help to better investigate the invasive behaviors of HCC cells.
