Study on Differentiation Potency Changes of Long-term Cultured Human Umbilical Cord MSCs

China Biotechnology

2012, Vol. 32

Issue (02): 8-16 DOI:

Study on Differentiation Potency Changes of Long-term Cultured Human Umbilical Cord MSCs

LIU Jing¹, YU Li¹, XU Chao², LUO Er-mei¹

1. Biochemistry Department, Jinan University, Guangzhou 510632, China;
2. Tin Ka Ping center for Medical Experiment, Jinan University, Guangzhou 510632, China

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Abstract

Objective: To investigate the ability of multilineage differentiation of human umbilical cord-MSCs by subculturing to 23 passages.Methods: Mesenchymal stem cells were isolated from umbilical cord using collagenaseⅡdigestion and were digested by trypsin for continuous passage culture. A series of different generation cultured-cells were collected for following experiment. The cells surface antigens and cell cycle were measured by flow cytometry; The cell growth curve was analyzed by MTT assay; Adipogenic、osteogenic and chondrogenetic ability were evaluated by specially inducing culture; OCT-4、SOX-2 and Nanog mRNA expression level were assayed by real-time fluorescence quantitative PCR. Result: MSCs can be obtained by collagenase digestion method. They had uniform shape, and can be stably passaged over 23 generations. By subculturing to 23 passages in vitro, the expression rate of surface marker in cell was not significant changed; the shape of cell growth curves were similarity; there was also no obvious difference in cell cycle, (73.04±1.15)% of the cell were G0/G1 phase; cells can be induced into osteoblasts, chondrocytes and adipocytes; OCT-4、SOX-2 and Nanog mRNA expression level were no obvious difference. Conclusion: Subculturing to 23 generation in vitro, the ability of mutilineage differentiation of hUC-MSCs was not influenced with increase in numbers of subculture.

Key words： Human umbilical cord-MSCs Long-term culture Multilineage differentiation Gene

Received: 27 July 2011 Published: 25 February 2012

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Cite this article:

LIU Jing, YU Li, XU Chao, LUO Er-mei. Study on Differentiation Potency Changes of Long-term Cultured Human Umbilical Cord MSCs. China Biotechnology, 2012, 32(02): 8-16.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2012/V32/I02/8

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