| 刘红王海涛李世崇叶玲玲刘兴茂陈昭烈.组成型过量表达hTERT 对Vero 细胞体外培养生物学特性的影响[J].,2011,11(11):2001-2005 |
| 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 Metabolismof Vero Cells Cultured in Vitro |
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| DOI： |
| 中文关键词: hTERT 过量表达 Vero 细胞 细胞培养 |
| 英文关键词: hTERT Overexpression Vero cells Cell culture |
| 基金项目:“重大新药创制”国家重大科技专项课题(2009ZX09503-011)，“十一五”国家科技支撑计划项目课题(2008BAI66B02) |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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 T1 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 T1 cells and wild-type Vero 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 of hTERT could reduce the dependency on serum and adhesion of
Vero cells cultured in vitro, which indicated that constitutive overexpression of hTERT is a potential technical approach to optimize the
characteristics of animal cells cultured in vitro. |
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