组成型过量表达hTERT对Vero细胞体外培养生物学特性的影响

Effects of Constitutively Overexpressing hTERT on Growth and Metabolism of Vero Cells Cultured in Vitro

目的:考察组成型过量表达人端粒酶催化亚单位(hTERT)对Vero细胞在无血清培养体系中的细胞形态、生长和代谢的影响。方法:以组成型过量表达hTERT的Vero细胞系T1为研究对象,以活细胞密度和细胞活力为主要观察指标,结合细胞形态和贴附伸展动态,考察T1细胞和野生型Vero细胞在静止贴附培养、微载体固定化培养和悬浮培养体系中的细胞生长;以葡萄糖比消耗速率(qglc)、乳酸比生成速率(qlac)、乳酸转化率(Ylac/glc)和谷氨酰胺比消耗率(qgln)为反映细胞代谢的主要观察指标,考察T1细胞和野生型Vero细胞在静止贴附培养、微载体固定化培养的细胞代谢。结果:hTERT组成型过量表达在降低Vero细胞的贴附伸展能力和对血清的依赖程度的同时,提高了细胞无血清批次培养后期的细胞活力和活细胞密度,并赋予了T1非贴附依赖性生长的能力。hTERT组成型过量表达未对Vero细胞的代谢产生明显的影响。结论:hTERT组成型过量表达可降低Vero细胞的贴附生长依赖性和对血清的依赖程度,是有应用潜力的改良哺乳动物细胞体外培养性状的技术途径。

Objective： To evaluate the influences of constitutively overexpressing human telomerase reverse transcriptase （hTERT） on the growth and metabolism of Vero cells cultured in vitro. Methods： Using the viable cell density, cell viability as the major evaluation indexes, in combination with cell morphology and stretching dynamics, the growth of both the hTERT-overexpressing Vero cell line TI and wild-type Vero cells cultured in static attachment and suspended microcarriers was evaluated; Using the specific consumption rate of glucose （qglc）, specific consumption rate of glutamine （qgln）, specific production rate of lactate （qlac） and yield of lactate to glucose （Ylac/glc） as the evaluation indexes, the metabolism of both TI cells and wild-type Veto cells cultured in static attachment and suspended microcarriers was determined. Results： Constitutively hTERT overexpression in T1 cells resulted in poorer capability of adhesion and extension, and reduced serum dependence, followed by the ability of adhesion independent growth in vitro to some extent. And no significant differences in metabolism between T1 cells and wild-type Vero cells cultured in static attachment and suspended microcarriers were observed. Conclusion： Constitutive overexpression ofhTERT could reduce the dependency on serum and adhesion of Veto cells cultured in vitro, which indicated that constitutive overexpression of hTERT is a potential technical approach to optimize the characteristics of animal ceils cultured in vitro.