Gupta Neelam, Taneja Ritu, Pandey Alok, Mukesh Manishi, Singh Hari, Gupta S C
Transgenic Research Laboratory, DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal 132001, Haryana, India.
Cell Biol Int. 2007 Oct;31(10):1257-64. doi: 10.1016/j.cellbi.2007.05.004. Epub 2007 May 21.
We assessed aging in continuous donor skin fibroblast cell line GGM5 up to the 25th passage by in vitro replicative senescence, telomere dynamics and chromosomal abnormalities. Cell proliferation rate increased from 0.84+/-0.26 (primary cells) to 1.20+/-0.17 (13-15 passage group) per day and reduced to 0.65+/-0.14 in 22-25 passages. Cell proliferation rate was reduced by 45.7% after 87.62 CPDs. Cell viability reduced from 100% to 97.4% up to the 25th passages. Frequency of beta gal(+) cells increased in successive passages and days in culture. The correlation coefficient between frequency of beta gal(+) cells and growth rate was -0.50 to -0.61. Loss of mean TRF length was 13.8 nucleotides (passage 15) to 95.4 nucleotides per cell division in later passages. All cells showed Robertsonian translocation in 22-25 passaged cells. The SCNT pre-implantation embryos production was highest (22.5%) in donor cells used from 10-15 passages as compared to early (<or=5) and late (22-25 passages). Our findings supports that cell proliferation rates, beta gal staining, mean TRF loss and karyological profile are useful marker for evaluation of competent nuclear donor.
我们通过体外复制性衰老、端粒动力学和染色体异常评估了连续传代至第25代的供体皮肤成纤维细胞系GGM5的衰老情况。细胞增殖率从每天0.84±0.26（原代细胞）增加到1.20±0.17（第13 - 15代组），并在第22 - 25代降至0.65±0.14。在87.62个群体倍增水平后，细胞增殖率降低了45.7%。到第25代时，细胞活力从100%降至97.4%。在连续传代和培养天数中，β - 半乳糖苷酶阳性（β gal(+)）细胞的频率增加。β gal(+)细胞频率与生长速率之间的相关系数为 - 0.50至 - 0.61。平均端粒限制片段（TRF）长度的损失在第15代时为13.8个核苷酸，在后期传代中每个细胞分裂为95.4个核苷酸。在第22 - 25代细胞中，所有细胞均显示罗伯逊易位。与早期（≤5代）和晚期（第22 - 25代）相比，使用第10 - 15代的供体细胞进行体细胞核移植（SCNT）的植入前胚胎产量最高（22.5%）。我们的数据支持细胞增殖率、β - 半乳糖苷酶染色、平均TRF损失和核型分析是评估合格核供体的有用指标。
