Adipose tissue-derived mesenchymal stem cell differentiation into cardiomyocyte-like cells after cryopreservation and passage*,**,☆

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Abstract

BACKGROUND:

Adipose tissue-derived mesenchymal stem cell is a kind of adult multipotential stem cells, and will become seed cells for cardiac tissue engineering and cell therapy of heart disease.

OBJECTIVE:

To observe adipose tissue-derived mesenchymal stem cell differentiation into cardiomyocyte-like cells after cryopreservation or passage for a long time.

DESIGN, TIME AND SETTING:

The controlled observational experiment was performed at the Tissue Engineering Laboratory in the Institute of Medical Equipment, Academy of Military Medical Sciences of Chinese PLA from September 2007 to June 2008.

MATERIALS:

Rat adipose tissue-derived mesenchymal stem cells were obtained from epididymis adipose pad of 2-month-old Wistar rats.

METHODS:

The third passage rat adipose tissue-derived mesenchymal stem cells were preserved in liquid nitrogen for six months and thawed, then the cells were induced with 10 μ mol/L 5-azacytidine. After 2 weeks, the mesenchymal stem cell differentiation potential was detected. Additionally, the third passage adipose tissue-derived mesenchymal stem cells were passed for long time in vitro, then induced with 10 μ mol/L 5-azacytidine. After 2 weeks, the cell differentiation potential was detected. The stem cells not induced with 5-azacytidine served as control group.

MAIN OUTCOME MEASURES:

Immumohistochemistrical staining was used to observe expression of α -sacromeric actin, α -skeletal myosin (only detected in skeletal muscle) and GATA-4.

REAULTS:

After cryopreservation, α -sacromeric actin and GATA-4 immumohistochemistrical staining were both positive in the induced adipose tissue-derived mesenchymal stem cells, the α -skeletal myosin staining was negative in the induced cells. After inducement, the immumohistochemistrical staining result in the passage 6 adipose tissue-derived mesenchymal stem cells was the same as the cells mentioned above. After inducement, in the passage 9 adipose tissue-derived mesenchymal stem cells, the immumohistochemistrical staining results of α -sacromeric actin, α -skeletal myosin, GATA-4 were all positive. Control groups were all negative.

CONCLUSION:

After cryopreservation, adipose tissue-derived mesenchymal stem cells could differentiate to cardiomyocyte-like cells. After passage for a long time, adipose tissue-derived mesenchymal stem cells could differentiate into cardiomyocyte-like cells, but could differentiate into skeletal muscle-like cells after induced with 5-azacytidine.

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