Stem cell-like memory T(TSCM)cells possess stem cell properties including multipotency and self-renewal and are being recognized as emerging players in various human diseases.Advanced technologies such as multiparamet...Stem cell-like memory T(TSCM)cells possess stem cell properties including multipotency and self-renewal and are being recognized as emerging players in various human diseases.Advanced technologies such as multiparametric flowcytometry and single cell sequencing have enabled their identification and molecular characterization.In case of chronic viral diseases such as human immunodeficiency virus-1,CD4+T_(SCM) cells,serve as major reservoirs of the latent virus.However,during immune activation and functional exhaustion of effector T cells,these cells also possess the potential to replenish the pool of functional effector cells to curtail the infection.More recently,these cells are speculated to play important role in protective immunity following acute viral infections such as coronavirus disease 2019 and might be amenable for therapeutics by ex vivo expansion.Similarly,studies are also investigating their pathological role in driving autoimmune responses.However,there are several gaps in the understanding of the role of T_(SCM) cells in viral and autoimmune diseases to make them potential therapeutic targets.In this minireview,we have attempted an updated compilation of the dyadic role of these complex T_(SCM) cells during such human diseases along with their biology and transcriptional programs.展开更多
BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural...BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural regeneration. OBJECTIVE： To explore the changes in S100 expression during rat MSCs differentiation into Schwann ceils in vitro. DESIGN, TIME AND SETTING： This cytology experiment was performed at the Jiangsu Key Laboratory of Neuroregeneration, Nantong University in China, from January 2006 to May 2007. MATERIALS： The rabbit anti-S100 polyclonal antibody was purchased from Dako, Denmark; the mouse anti-rat S100 monoclonal antibody was purchased from Sigma, USA. METHODS： MSCs were cultured from adult Sprague-Dawley rat femur and tibia. Cell proliferation was determined by the MTT method and CD markers, and cell cycle was measured by flow cytometry. MSCs were induced to differentiate into SC cells. SC cells were stained for S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor. S100 protein and mRNA levels were evaluated by flow cytometry, Western blot, and reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURES： S100 protein and mRNA expression. RESULTS： MSCs exhibited high amplification potential over eight passages. Prior to induction, the majority of MSCs were at the G0/G1 phase of the cell cycle. After induction, MSCs displayed morphology changes similar to Schwann cells. Moreover, induction increased S100 mRNA levels. Immunofluorescence showed that MSCs expressed S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor at 7 days of induction. Induction also increased S100 protein levels compared with untreated MSCs. CONCLUSION： MSCs are capable of differentiating into Schwann cells-like cells under conditional induction in vitro, with increasing S100 mRNA and protein expression.展开更多
Prostate cancers(PCa)have been reported to actively suppress antitumor immune responses by creating an immune-suppressive microenvironment.There is mounting evidence that PCas may undergo an‘‘Epithelial Immune Cell-...Prostate cancers(PCa)have been reported to actively suppress antitumor immune responses by creating an immune-suppressive microenvironment.There is mounting evidence that PCas may undergo an‘‘Epithelial Immune Cell-like Transition’’(EIT)by expressing molecules conventionally associated with immune cells(e.g.,a variety of cytokines/receptors,immune transcription factors,Ig motifs,and immune checkpoint molecules),which subsequently results in the suppression of anti-cancer immune activity within the tumor microenvironment.Recent progress within the field of immune therapy has underscored the importance of immune checkpoint molecules in cancer development,thus leading to the development of novel immunotherapeutic approaches.Here,we review the expression of select immune checkpoint molecules in PCa epithelial and associated immune cells,with particular emphasis on clinical data supporting the concept of an EIT-mediated phenotype in PCa.Furthermore,we summarize current advances in anti-immune checkpoint therapies,and provide perspectives on their potential applicability.展开更多
Objective： Colon cancer stem cells （CSCs） are implicated in colorectal cancer carcinogenesis, metastasis, and therapeutic resistance. The identification of these cells could help to develop novel therapeutic strate...Objective： Colon cancer stem cells （CSCs） are implicated in colorectal cancer carcinogenesis, metastasis, and therapeutic resistance. The identification of these cells could help to develop novel therapeutic strategies. Doublecortin-like kinase 1 （DCLK1） has been viewed as a marker for gastrointestinal stem cells that fuel the self-renewal process, however others view them as a marker of Tuft cells or as an enteroendocrine subtype. The purpose of this study was to use a colon cancer cell line to identify and characterize the stem-like characteristics of the DCLKI＋ cell population. Methods： To enrich stem-like cells, HCT116 cells （derived from colon adenocarcinomas） were cultured using serum-free media to form spheres under both normal oxygen and hypoxia condition. DCLK1 transcript expression in the adherent parental cells and spheroids was quantified using quantitative real time reverse transcription- polymerase chain reaction [（q）RT-PCR]. DCLK1 protein expression was determined using flow cytometry. Self-renewal capability from adherent parental cells and spheroids was determined using extreme limiting dilution analysis （ELDA）. Results： Under both normal oxygen and hypoxia condition, the adherent parental cells were composed of cells that express low levels of DCLK1. However, spheroids exhibited an increased frequency of cells expressing DCLK1 on both mRNA and protein levels. Cells derived from spheroids also possess stronger self-renewal capability. Conclusions： The higher fraction of DCLK1 ＋ cells exhibited by spheroids and hypoxia reflects the stem- like characteristics of these cells. DCLK1 may represent an ideal marker to study and develop effective strategies to overcome chemo-resistance and relapse of colon cancer.展开更多
The cancer stem cell hypothesis provides a basis for prediction of the recurrence and risk of metastasis in breast cancer.However,the unique expression pattern of stemness markers and the presence of nonstem-like canc...The cancer stem cell hypothesis provides a basis for prediction of the recurrence and risk of metastasis in breast cancer.However,the unique expression pattern of stemness markers and the presence of nonstem-like cancer cells with varied phenotypes have brought great challenges to the characterization of breast cancer stem cells.To address these challenges,a phenotype-directed DNA nanomachine has been designed for high-accuracy labeling and in situ analysis of the stem cell-like subpopulation in breast cancer.The key for the design is to use cell surfaceanchored inputs to activate the nanomachine,which undergoes different branch migration pathways such that the signal strand can only be brought onto the cancer cells having the stem cell-like phenotype.Highly sensitive determination and single-step isolation of the stem cell-like subpopulation were achieved by incorporating functional groups into the signal strand such that the nanomachine was successfully applied in a tumor-bearing mouse model.Overall,the approach provides for a substantial improvement in capability for the analysis of the breast cancer stem cell-like subpopulation,and it is expected that the new approach will advance the use of DNA nanomachines in cancer-related studies.展开更多
Background:Schwann cell-like cells(SCLCs),differentiated from mesenchymal stem cells,have shown promising outcomes in the treatment of peripheral nerve injuries in preclinical studies.However,certain clinical obstacle...Background:Schwann cell-like cells(SCLCs),differentiated from mesenchymal stem cells,have shown promising outcomes in the treatment of peripheral nerve injuries in preclinical studies.However,certain clinical obstacles limit their application.Hence,the primary aim of this study was to investigate the role of exosomes derived from SCLCs(SCLCs-exo)in peripheral nerve regeneration.Methods:SCLCs were differentiated from human amniotic mesenchymal stem cells(hAMSCs)in vitro and validated by immunofluorescence,real-time quantitative PCR and western blot analysis.Exosomes derived from hAMSCs(hAMSCs-exo)and SCLCs were isolated by ultracentrifugation and validated by nanoparticle tracking analysis,WB analysis and electron microscopy.A prefab-ricated nerve graft was used to deliver hAMSCs-exo or SCLCs-exo in an injured sciatic nerve rat model.The effects of hAMSCs-exo or SCLCs-exo on rat peripheral nerve injury(PNI)regeneration were determined based on the recovery of neurological function and histomorphometric variation.The effects of hAMSCs-exo or SCLCs-exo on Schwann cells were also determined via cell prolifer-ation and migration assessment.Results:SCLCs significantly expressed the Schwann cell markers glial fibrillary acidic protein and S100.Compared to hAMSCs-exo,SCLCs-exo significantly enhanced motor function recov-ery,attenuated gastrocnemius muscle atrophy and facilitated axonal regrowth,myelin forma-tion and angiogenesis in the rat model.Furthermore,hAMSCs-exo and SCLCs-exo were effi-ciently absorbed by Schwann cells.However,compared to hAMSCs-exo,SCLCs-exo signifi-cantly promoted the proliferation and migration of Schwann cells.SCLCs-exo also significantly upregulated the expression of a glial cell-derived neurotrophic factor,myelin positive regulators(SRY-box transcription factor 10,early growth response protein 2 and organic cation/carnitine transporter 6)and myelin proteins(myelin basic protein and myelin protein zero)in Schwann cells.Conclusions:These findings suggest that SCLCs-exo can more efficiently promote PNI regeneration than hAMSCs-exo and are a potentially novel therapeutic approach for treating PNI.展开更多
Bio-mimicking graphene films,deposited on textured nickel substrates,were synthesized by the following method:replicating the surface textures of the lotus leaf by polymer duplication,fabricating textured nickel subst...Bio-mimicking graphene films,deposited on textured nickel substrates,were synthesized by the following method:replicating the surface textures of the lotus leaf by polymer duplication,fabricating textured nickel substrates by electroplating on the polymer coated with a Au film,preparing bio-mimicking graphene oxide films on the nickel substrates by vacuum filtration,and electrochemical reduction.By controlling the vacuum filtration,this replica method can not only replicate the lotus leaf structure by a graphene film,but also can achieve a novel cell-like graphene film.展开更多
Basic magnesium carbonate microspheres with a red blood cell （RBC）-like appearance and diameters of ~3μm were synthesized by amphiphilic molecule-participated self-assembly under hydrothermal conditions, In the sel...Basic magnesium carbonate microspheres with a red blood cell （RBC）-like appearance and diameters of ~3μm were synthesized by amphiphilic molecule-participated self-assembly under hydrothermal conditions, In the self-assembly, sodium dodecyl benzene sulfonate served as a template for the formation of Mg（OH）2 spherical micelles and also as a reactant precursor that releases CO2 to react with Mg（OH）2. The growth of the microspheres is driven by the continuous generation of new hydrophobic centers because of the consumption of hydrophilic poles （--SO3-）. The surfactant-directed self-assembly can be applied to the synthesis of other carbonate or metallic oxide self-assemblies, indicating that it is a universal self-assembly method for amphiphilic molecules.展开更多
During the last decade,increasing evidence suggested that bone marrow stromal cells(MSCs) have the potential to differentiate into neural lineages.Many studies have reported that MSCs showed morphological changes and ...During the last decade,increasing evidence suggested that bone marrow stromal cells(MSCs) have the potential to differentiate into neural lineages.Many studies have reported that MSCs showed morphological changes and expressed a limited number of neural proteins under experimental conditions.However,no proteomic studies on MSCs differentiated into Schwann cell-like cells have been reported.In this study,we isolated MSCs from adult Sprague-Dawley rat femur and tibia bone marrows and induced the cells in vitro under specific conditions.By using two-dimensional gel electrophoresis(2-DE),we compared the protein profiles of MSCs before and after induced differentiation.We obtained 792 protein spots in the protein profile by 2-DE,and found that 74 spots changed significantly before and after the differentiation using PDQuest software,with 43 up-regulated and 31 down-regulated.We analyzed these 74 spots by a matrix assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS) and by database searching,and found that they could be grouped into various classes,including cytoskeleton and structure proteins,growth factors,metabolic proteins,chaperone proteins,receptor proteins,cell cycle proteins,calcium binding proteins,and other proteins.These proteins also include neural and glial proteins,such as BDNF,CNTF and GFAP.The results may provide valuable proteomic information about the differentiation of MSCs into Schwann cell-like cells.展开更多
文摘Stem cell-like memory T(TSCM)cells possess stem cell properties including multipotency and self-renewal and are being recognized as emerging players in various human diseases.Advanced technologies such as multiparametric flowcytometry and single cell sequencing have enabled their identification and molecular characterization.In case of chronic viral diseases such as human immunodeficiency virus-1,CD4+T_(SCM) cells,serve as major reservoirs of the latent virus.However,during immune activation and functional exhaustion of effector T cells,these cells also possess the potential to replenish the pool of functional effector cells to curtail the infection.More recently,these cells are speculated to play important role in protective immunity following acute viral infections such as coronavirus disease 2019 and might be amenable for therapeutics by ex vivo expansion.Similarly,studies are also investigating their pathological role in driving autoimmune responses.However,there are several gaps in the understanding of the role of T_(SCM) cells in viral and autoimmune diseases to make them potential therapeutic targets.In this minireview,we have attempted an updated compilation of the dyadic role of these complex T_(SCM) cells during such human diseases along with their biology and transcriptional programs.
基金the National High-Tech Research & Development Program of China, No. 2006AA02A128the National Natural Science Foundation of China, No. 30670667
文摘BACKGROUND： S100 protein can promote axonal growth. Therefore, transplantation of induced bone marrow-derived mesenchymal stem cells （MSCs） that can secrete S100 may provide a beneficial microenvironment for neural regeneration. OBJECTIVE： To explore the changes in S100 expression during rat MSCs differentiation into Schwann ceils in vitro. DESIGN, TIME AND SETTING： This cytology experiment was performed at the Jiangsu Key Laboratory of Neuroregeneration, Nantong University in China, from January 2006 to May 2007. MATERIALS： The rabbit anti-S100 polyclonal antibody was purchased from Dako, Denmark; the mouse anti-rat S100 monoclonal antibody was purchased from Sigma, USA. METHODS： MSCs were cultured from adult Sprague-Dawley rat femur and tibia. Cell proliferation was determined by the MTT method and CD markers, and cell cycle was measured by flow cytometry. MSCs were induced to differentiate into SC cells. SC cells were stained for S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor. S100 protein and mRNA levels were evaluated by flow cytometry, Western blot, and reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURES： S100 protein and mRNA expression. RESULTS： MSCs exhibited high amplification potential over eight passages. Prior to induction, the majority of MSCs were at the G0/G1 phase of the cell cycle. After induction, MSCs displayed morphology changes similar to Schwann cells. Moreover, induction increased S100 mRNA levels. Immunofluorescence showed that MSCs expressed S100 protein, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor at 7 days of induction. Induction also increased S100 protein levels compared with untreated MSCs. CONCLUSION： MSCs are capable of differentiating into Schwann cells-like cells under conditional induction in vitro, with increasing S100 mRNA and protein expression.
文摘Prostate cancers(PCa)have been reported to actively suppress antitumor immune responses by creating an immune-suppressive microenvironment.There is mounting evidence that PCas may undergo an‘‘Epithelial Immune Cell-like Transition’’(EIT)by expressing molecules conventionally associated with immune cells(e.g.,a variety of cytokines/receptors,immune transcription factors,Ig motifs,and immune checkpoint molecules),which subsequently results in the suppression of anti-cancer immune activity within the tumor microenvironment.Recent progress within the field of immune therapy has underscored the importance of immune checkpoint molecules in cancer development,thus leading to the development of novel immunotherapeutic approaches.Here,we review the expression of select immune checkpoint molecules in PCa epithelial and associated immune cells,with particular emphasis on clinical data supporting the concept of an EIT-mediated phenotype in PCa.Furthermore,we summarize current advances in anti-immune checkpoint therapies,and provide perspectives on their potential applicability.
文摘Objective： Colon cancer stem cells （CSCs） are implicated in colorectal cancer carcinogenesis, metastasis, and therapeutic resistance. The identification of these cells could help to develop novel therapeutic strategies. Doublecortin-like kinase 1 （DCLK1） has been viewed as a marker for gastrointestinal stem cells that fuel the self-renewal process, however others view them as a marker of Tuft cells or as an enteroendocrine subtype. The purpose of this study was to use a colon cancer cell line to identify and characterize the stem-like characteristics of the DCLKI＋ cell population. Methods： To enrich stem-like cells, HCT116 cells （derived from colon adenocarcinomas） were cultured using serum-free media to form spheres under both normal oxygen and hypoxia condition. DCLK1 transcript expression in the adherent parental cells and spheroids was quantified using quantitative real time reverse transcription- polymerase chain reaction [（q）RT-PCR]. DCLK1 protein expression was determined using flow cytometry. Self-renewal capability from adherent parental cells and spheroids was determined using extreme limiting dilution analysis （ELDA）. Results： Under both normal oxygen and hypoxia condition, the adherent parental cells were composed of cells that express low levels of DCLK1. However, spheroids exhibited an increased frequency of cells expressing DCLK1 on both mRNA and protein levels. Cells derived from spheroids also possess stronger self-renewal capability. Conclusions： The higher fraction of DCLK1 ＋ cells exhibited by spheroids and hypoxia reflects the stem- like characteristics of these cells. DCLK1 may represent an ideal marker to study and develop effective strategies to overcome chemo-resistance and relapse of colon cancer.
基金the National Natural Science Foundation of China(grant nos.81972799 and 81871449)the Natural Science Foundation of Shanghai(grant no.23ZR1421400).
文摘The cancer stem cell hypothesis provides a basis for prediction of the recurrence and risk of metastasis in breast cancer.However,the unique expression pattern of stemness markers and the presence of nonstem-like cancer cells with varied phenotypes have brought great challenges to the characterization of breast cancer stem cells.To address these challenges,a phenotype-directed DNA nanomachine has been designed for high-accuracy labeling and in situ analysis of the stem cell-like subpopulation in breast cancer.The key for the design is to use cell surfaceanchored inputs to activate the nanomachine,which undergoes different branch migration pathways such that the signal strand can only be brought onto the cancer cells having the stem cell-like phenotype.Highly sensitive determination and single-step isolation of the stem cell-like subpopulation were achieved by incorporating functional groups into the signal strand such that the nanomachine was successfully applied in a tumor-bearing mouse model.Overall,the approach provides for a substantial improvement in capability for the analysis of the breast cancer stem cell-like subpopulation,and it is expected that the new approach will advance the use of DNA nanomachines in cancer-related studies.
基金supported by the InnovationGroup Major Research Project of Guizhou Province Education Department(No.Qianjiaohe KY[2017]043)the Science and Technology Support Project of Guizhou Province(2020-5012)+3 种基金the PhD Fund of Scientific Research Foundation of the Affiliated Hospital of ZunyiMedical University(2020-03)the National Nature Science Foundation of China(81660325)the Collaborative Innovation Center of the Chinese Ministry of Education(2020-39)the Master Fund of Scientific Research Foundation of the Affiliated Hospital of Zunyi Medical University(2016-35).
文摘Background:Schwann cell-like cells(SCLCs),differentiated from mesenchymal stem cells,have shown promising outcomes in the treatment of peripheral nerve injuries in preclinical studies.However,certain clinical obstacles limit their application.Hence,the primary aim of this study was to investigate the role of exosomes derived from SCLCs(SCLCs-exo)in peripheral nerve regeneration.Methods:SCLCs were differentiated from human amniotic mesenchymal stem cells(hAMSCs)in vitro and validated by immunofluorescence,real-time quantitative PCR and western blot analysis.Exosomes derived from hAMSCs(hAMSCs-exo)and SCLCs were isolated by ultracentrifugation and validated by nanoparticle tracking analysis,WB analysis and electron microscopy.A prefab-ricated nerve graft was used to deliver hAMSCs-exo or SCLCs-exo in an injured sciatic nerve rat model.The effects of hAMSCs-exo or SCLCs-exo on rat peripheral nerve injury(PNI)regeneration were determined based on the recovery of neurological function and histomorphometric variation.The effects of hAMSCs-exo or SCLCs-exo on Schwann cells were also determined via cell prolifer-ation and migration assessment.Results:SCLCs significantly expressed the Schwann cell markers glial fibrillary acidic protein and S100.Compared to hAMSCs-exo,SCLCs-exo significantly enhanced motor function recov-ery,attenuated gastrocnemius muscle atrophy and facilitated axonal regrowth,myelin forma-tion and angiogenesis in the rat model.Furthermore,hAMSCs-exo and SCLCs-exo were effi-ciently absorbed by Schwann cells.However,compared to hAMSCs-exo,SCLCs-exo signifi-cantly promoted the proliferation and migration of Schwann cells.SCLCs-exo also significantly upregulated the expression of a glial cell-derived neurotrophic factor,myelin positive regulators(SRY-box transcription factor 10,early growth response protein 2 and organic cation/carnitine transporter 6)and myelin proteins(myelin basic protein and myelin protein zero)in Schwann cells.Conclusions:These findings suggest that SCLCs-exo can more efficiently promote PNI regeneration than hAMSCs-exo and are a potentially novel therapeutic approach for treating PNI.
基金supported by the"Hundred Talants Program"of the Chinese Academy of Sciences and the National Natural Science Foundation of China(51005225and51002161)
文摘Bio-mimicking graphene films,deposited on textured nickel substrates,were synthesized by the following method:replicating the surface textures of the lotus leaf by polymer duplication,fabricating textured nickel substrates by electroplating on the polymer coated with a Au film,preparing bio-mimicking graphene oxide films on the nickel substrates by vacuum filtration,and electrochemical reduction.By controlling the vacuum filtration,this replica method can not only replicate the lotus leaf structure by a graphene film,but also can achieve a novel cell-like graphene film.
基金supported by the National Natural Science Foundation of China(No.21206191)the Science Foundation of China University of Petroleum,Beijing(No.2462013YXBS007)
文摘Basic magnesium carbonate microspheres with a red blood cell （RBC）-like appearance and diameters of ~3μm were synthesized by amphiphilic molecule-participated self-assembly under hydrothermal conditions, In the self-assembly, sodium dodecyl benzene sulfonate served as a template for the formation of Mg（OH）2 spherical micelles and also as a reactant precursor that releases CO2 to react with Mg（OH）2. The growth of the microspheres is driven by the continuous generation of new hydrophobic centers because of the consumption of hydrophilic poles （--SO3-）. The surfactant-directed self-assembly can be applied to the synthesis of other carbonate or metallic oxide self-assemblies, indicating that it is a universal self-assembly method for amphiphilic molecules.
基金Supported by National High-Tech Research and Development Program of China(Grant No.2006AA02A128)National Natural Science Foundation of China(Grant No.30670667)
文摘During the last decade,increasing evidence suggested that bone marrow stromal cells(MSCs) have the potential to differentiate into neural lineages.Many studies have reported that MSCs showed morphological changes and expressed a limited number of neural proteins under experimental conditions.However,no proteomic studies on MSCs differentiated into Schwann cell-like cells have been reported.In this study,we isolated MSCs from adult Sprague-Dawley rat femur and tibia bone marrows and induced the cells in vitro under specific conditions.By using two-dimensional gel electrophoresis(2-DE),we compared the protein profiles of MSCs before and after induced differentiation.We obtained 792 protein spots in the protein profile by 2-DE,and found that 74 spots changed significantly before and after the differentiation using PDQuest software,with 43 up-regulated and 31 down-regulated.We analyzed these 74 spots by a matrix assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS) and by database searching,and found that they could be grouped into various classes,including cytoskeleton and structure proteins,growth factors,metabolic proteins,chaperone proteins,receptor proteins,cell cycle proteins,calcium binding proteins,and other proteins.These proteins also include neural and glial proteins,such as BDNF,CNTF and GFAP.The results may provide valuable proteomic information about the differentiation of MSCs into Schwann cell-like cells.
