Cell lines with high passage numbers exhibit alterations in cell morphology and functions.In thepresent work,C2C12 skeletal muscle cells with either low(<20)or high(>60)passage numbers(identified as l-C2C12 or h...Cell lines with high passage numbers exhibit alterations in cell morphology and functions.In thepresent work,C2C12 skeletal muscle cells with either low(<20)or high(>60)passage numbers(identified as l-C2C12 or h-C2C12,respectively)were used to investigate the apoptotic response to H2O2 as a function ofculture age h-C2C12.We found that older cultures(h-C2C12 group)were depleted of mitochondrial DNA(mtDNA).When we analyzed the behavior of Bad,Bax,caspase-3 and mitochondrial transmembrane potential,we observed that cells in the h-C2C12 group were resistant to H2O2 induction of apoptosis.We propose seriallycultured C2C12 cells as a refractory model to H2O2-induced apoptosis.In addition,the data obtained in thiswork suggest that mtDNA is required for apoptotic cell death in skeletal muscle C2C12 cells.展开更多
To investigate the biological character of human adipose-derived adult stem cells (hADAS cells) when cultured in vitro and the relationship between hADAS cell’s replication activity and the donor’s age factor, and t...To investigate the biological character of human adipose-derived adult stem cells (hADAS cells) when cultured in vitro and the relationship between hADAS cell’s replication activity and the donor’s age factor, and to assess the stem cells as a new source for tissue engineering. hADAS cells are isolated from human adipose tissue of different age groups (from adolescents to olds: <20 years old, 21―40 years old, 41―60 years old and >61 years old groups). The protein markers (CD29, CD34, CD44, CD45, CD49d, HLA-DR, CD106) of hADAS cells were detected by flow cytometry (FCM) to identify the stem cell, and the cell cycle was examined for P20 hADAS cells to evaluate the safety of the subculture in vitro. The generative activity of hADAS cells in different age groups was also examined by MTT method. The formula “ log2T D = t logN t ? logN 0” was used to get the time doubling (TD) of the cells. The results showed that the cells kept heredity stabilization by chromosome analysis for at least 20 passages. The TD of these cells increased progressively by ageing, and the TD of the <20 years old group was lower than that of the >61 years old group (statistical analysis of variance (ANOVA), P=0.002, P<0.05). These find- ings suggested that a higher level of hADAS cells replication activity was found in the younger dona- tors, and they represent novel and valuable seed cells for studies of tissue engineering.展开更多
基金supported by the NationalResearch Council of Argentina(CONICET)and theUniversidad Nacional del Sur,Argentina.
文摘Cell lines with high passage numbers exhibit alterations in cell morphology and functions.In thepresent work,C2C12 skeletal muscle cells with either low(<20)or high(>60)passage numbers(identified as l-C2C12 or h-C2C12,respectively)were used to investigate the apoptotic response to H2O2 as a function ofculture age h-C2C12.We found that older cultures(h-C2C12 group)were depleted of mitochondrial DNA(mtDNA).When we analyzed the behavior of Bad,Bax,caspase-3 and mitochondrial transmembrane potential,we observed that cells in the h-C2C12 group were resistant to H2O2 induction of apoptosis.We propose seriallycultured C2C12 cells as a refractory model to H2O2-induced apoptosis.In addition,the data obtained in thiswork suggest that mtDNA is required for apoptotic cell death in skeletal muscle C2C12 cells.
基金the Science and Technical Research Funds of Guangdong Province, China (Grant Nos. 2004B34001004 and04009423)
文摘To investigate the biological character of human adipose-derived adult stem cells (hADAS cells) when cultured in vitro and the relationship between hADAS cell’s replication activity and the donor’s age factor, and to assess the stem cells as a new source for tissue engineering. hADAS cells are isolated from human adipose tissue of different age groups (from adolescents to olds: <20 years old, 21―40 years old, 41―60 years old and >61 years old groups). The protein markers (CD29, CD34, CD44, CD45, CD49d, HLA-DR, CD106) of hADAS cells were detected by flow cytometry (FCM) to identify the stem cell, and the cell cycle was examined for P20 hADAS cells to evaluate the safety of the subculture in vitro. The generative activity of hADAS cells in different age groups was also examined by MTT method. The formula “ log2T D = t logN t ? logN 0” was used to get the time doubling (TD) of the cells. The results showed that the cells kept heredity stabilization by chromosome analysis for at least 20 passages. The TD of these cells increased progressively by ageing, and the TD of the <20 years old group was lower than that of the >61 years old group (statistical analysis of variance (ANOVA), P=0.002, P<0.05). These find- ings suggested that a higher level of hADAS cells replication activity was found in the younger dona- tors, and they represent novel and valuable seed cells for studies of tissue engineering.
