BACKGROUND Human adipose-derived stromal/stem cells(hASCs)are one of the most useful types of mesenchymal stromal/stem cells,which are adult multipotent cells with great therapeutic potential for the treatment of seve...BACKGROUND Human adipose-derived stromal/stem cells(hASCs)are one of the most useful types of mesenchymal stromal/stem cells,which are adult multipotent cells with great therapeutic potential for the treatment of several diseases.However,for successful clinical application,it is critical that high-quality cells can be obtained.Diverse factors seem to be able to influence cell quality and performance,especially factors related to donors’intrinsic characteristics,such as age.Nevertheless,there is no consensus regarding this characteristic,and there is conflicting information in the literature.AIM To investigate the growth kinetics and differentiation potential of adipose-derived stem cells isolated from the lipoaspirates of elderly and young donors.METHODS hASCs were harvested from liposuctioned adipose tissue obtained from female donors(aged 20-70 years).Cells were distributed into two groups according to age range:old hASCs(oASCs,≥55 years,n=9)and young hASCs(yASCs,≤35 years,n=9).For each group,immunophenotypic characterization was performed by flow cytometry.Population doubling time was assessed over seven days.For adipogenic potential evaluation,lipid deposits were assessed after 7 d,14 d and 21 d of adipogenic induction.Osteogenic potential was verified by analyzing cell mineralization after 14 d,21 d and 28 d of osteogenic induction.mRNA expression of PPARγ2,CEBPA and Runx2 were detected by quantitative reverse transcription polymerase chain reaction.RESULTS hASCs were successfully obtained,cultured,and grouped according to their age:yASCs(26.33±4.66 years old)and oASCs(64.78±4.58 years old).After maintenance of the cells in culture,there were no differences in morphology between cells from the young and old donors.Additionally,both groups showed classical immunophenotypic characteristics of mesenchymal stem/stromal cells.The average doubling time indicated that yASCs(4.09±0.94 d)did not significantly differ from oASCs(4.19±1.29 d).Concerning differentiation potential,after adipogenic and osteogenic induction,yASCs and oASCs were able to differentiate to greater levels than the noninduced control cells.However,no differences were found in the differentiation efficiency of yASCs and oASCs in adipogenesis or osteogenesis.Additionally,the mRNA expression of PPARγ2,CEBPA and Runx2 were similar in yASCs and oASCs.CONCLUSION Our findings suggest that age does not seem to significantly affect the cell division or adipogenic or osteogenic differentiation ability of adipose-derived stem cells isolated from lipoaspirates.展开更多
Tissue-specific resident cells have been identified as a promising population of progenitor cells for cell-based therapies. We describe here the isolation from adult human hearts of tissue nonspecific alkaline phospha...Tissue-specific resident cells have been identified as a promising population of progenitor cells for cell-based therapies. We describe here the isolation from adult human hearts of tissue nonspecific alkaline phosphatase-positive cells (ALPL+ cells) with mesenchymal stem cell (MSC) characteristics. Samples from 24 adult cadaveric donors were obtained from a valve bank. Mean total ischemia time was 21.5 ± 9.1 hours. The success rate for the isolation of human heart-derived cells by the explant culture technique was 70% for the right auricle (14 of 20 trials) and 33% for the right ventricle (7 of 21 trials). The total auricle-derived cell population (TAD) was used for the purification of ALPL+ cells. TAD and ALPL+ cells expressed markers for MSC and pericytes. TAD cells and ALPL+ cells differentiated into adipocytes, osteoblasts and chondroblasts, and ALPL+ cells expressed markers of these three lineages more strongly than TAD cells, as shown by RT-PCR. This population therefore has a greater potential for differentiation into mesechymal lineages than TAD cells. Both cell populations express some markers of cardiac progenitors, such as GATA4, CD117 and VEGF. ALPL+ cells expressed troponin T and ABCG2, which are also markers of the cardiac lineage. Heart samples from tissue banks could be considered as sources of MSC with putative commitment towards cardiac lineages, even after prolonged ischemia times.展开更多
基金Supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico,No.442353/2019-7 and No.442375/2019-0.
文摘BACKGROUND Human adipose-derived stromal/stem cells(hASCs)are one of the most useful types of mesenchymal stromal/stem cells,which are adult multipotent cells with great therapeutic potential for the treatment of several diseases.However,for successful clinical application,it is critical that high-quality cells can be obtained.Diverse factors seem to be able to influence cell quality and performance,especially factors related to donors’intrinsic characteristics,such as age.Nevertheless,there is no consensus regarding this characteristic,and there is conflicting information in the literature.AIM To investigate the growth kinetics and differentiation potential of adipose-derived stem cells isolated from the lipoaspirates of elderly and young donors.METHODS hASCs were harvested from liposuctioned adipose tissue obtained from female donors(aged 20-70 years).Cells were distributed into two groups according to age range:old hASCs(oASCs,≥55 years,n=9)and young hASCs(yASCs,≤35 years,n=9).For each group,immunophenotypic characterization was performed by flow cytometry.Population doubling time was assessed over seven days.For adipogenic potential evaluation,lipid deposits were assessed after 7 d,14 d and 21 d of adipogenic induction.Osteogenic potential was verified by analyzing cell mineralization after 14 d,21 d and 28 d of osteogenic induction.mRNA expression of PPARγ2,CEBPA and Runx2 were detected by quantitative reverse transcription polymerase chain reaction.RESULTS hASCs were successfully obtained,cultured,and grouped according to their age:yASCs(26.33±4.66 years old)and oASCs(64.78±4.58 years old).After maintenance of the cells in culture,there were no differences in morphology between cells from the young and old donors.Additionally,both groups showed classical immunophenotypic characteristics of mesenchymal stem/stromal cells.The average doubling time indicated that yASCs(4.09±0.94 d)did not significantly differ from oASCs(4.19±1.29 d).Concerning differentiation potential,after adipogenic and osteogenic induction,yASCs and oASCs were able to differentiate to greater levels than the noninduced control cells.However,no differences were found in the differentiation efficiency of yASCs and oASCs in adipogenesis or osteogenesis.Additionally,the mRNA expression of PPARγ2,CEBPA and Runx2 were similar in yASCs and oASCs.CONCLUSION Our findings suggest that age does not seem to significantly affect the cell division or adipogenic or osteogenic differentiation ability of adipose-derived stem cells isolated from lipoaspirates.
文摘Tissue-specific resident cells have been identified as a promising population of progenitor cells for cell-based therapies. We describe here the isolation from adult human hearts of tissue nonspecific alkaline phosphatase-positive cells (ALPL+ cells) with mesenchymal stem cell (MSC) characteristics. Samples from 24 adult cadaveric donors were obtained from a valve bank. Mean total ischemia time was 21.5 ± 9.1 hours. The success rate for the isolation of human heart-derived cells by the explant culture technique was 70% for the right auricle (14 of 20 trials) and 33% for the right ventricle (7 of 21 trials). The total auricle-derived cell population (TAD) was used for the purification of ALPL+ cells. TAD and ALPL+ cells expressed markers for MSC and pericytes. TAD cells and ALPL+ cells differentiated into adipocytes, osteoblasts and chondroblasts, and ALPL+ cells expressed markers of these three lineages more strongly than TAD cells, as shown by RT-PCR. This population therefore has a greater potential for differentiation into mesechymal lineages than TAD cells. Both cell populations express some markers of cardiac progenitors, such as GATA4, CD117 and VEGF. ALPL+ cells expressed troponin T and ABCG2, which are also markers of the cardiac lineage. Heart samples from tissue banks could be considered as sources of MSC with putative commitment towards cardiac lineages, even after prolonged ischemia times.
