Chondrogenic differentiation of co-cultured human umbilical cord blood-derived mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.2013.23.003

Abstract

Abstract:

BACKGROUND: Human umbilical cord blood-derived mesenchymal stem cells can be induced through the co-culture to differentiate into other cells; however, it is difficult to control the proportions of two kinds of cells for maximizing the efficiency of obtaining target cells.
OBJECTIVE: To investigate chondrogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells by co-culture with rabbit chondrocytes at different ratios.
METHODS: Human umbilical cord blood-derived mesenchymal stem cells were resuscitated and cultured, and identified by flow cytometry. Then human umbilical cord blood-derived mesenchymal stem cells were co-cultured with rabbit chondrocytes at 2:1 or 3:1 ratio, and induced with insulin-like growth factor-1. After 14 days in co-culture, the cell RNA and protein were extracted, aggrecan mRNA and collagen type Ⅱ mRNA expression levels were detected by real-time quantitative-polymerase chain reaction, and aggrecan and collagen type Ⅱ protein expression levels were detected by western blot assay.
RESULTS AND CONCLUSION: After human umbilical cord blood-derived mesenchymal stem cells were co-cultured with rabbit chondrocytes for 14 days, the aggrecan mRNA and collagen type 2 mRNA expression in the co-cultured cells was significantly higher than that in the cells only induced with insulin-like growth factor-1. Under the same ratio of human umbilical cord blood-derived mesenchymal stem cells and rabbit chondrocytes, insulin-like growth factor-1 increased the aggrecan and collagen type Ⅱ protein and mRNA expressions. A 3:1 co-culture ratio allowed the increment in the collagen type Ⅱ mRNA and protein expression, as well as aggrecan protein expression; while a 2:1 co-culture ratio upregulated aggrecan mRNA expression. Experimental findings indicate that, human chondrocytes can be successfully induced by co-culture of human umbilical cord blood-derived mesenchymal stem cells and rabbit chondrocytes, and 3:1 co-culture ratio can obtain more chondrocytes.

Key words: stem cells, umbilical cord/umbilical cord blood stem cells, chondrocytes, co-culture, insulin-like growth factor-1, collagen type Ⅱ, aggrecan, cartilage defects, cartilage tissue engineering, seed cells, other grants-supported paper

CLC Number:

R394.2

Zheng Peng-fei, Chen Lei, Dong Zhan, Jiang Li, Ju Li, Wang Ru-fa, Lou Yue. Chondrogenic differentiation of co-cultured human umbilical cord blood-derived mesenchymal stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(23): 4196-4203.

Figures/Tables 4

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