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High efficient isolation and systematic identification of human adipose-derived mesenchymal stem cells

Xu-Fang Yang1,2, Xu He1*, Jian He1, Li-Hong Zhang1, Xue-Jin Su1, Zhi-Yong Dong1, Yun-Jian Xu3, Yan Li4 and Yu-Lin Li1*

Author Affiliations

1 Key Laboratory of Pathobiology, Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, China

2 Department of Pathophysiology, MuDanJiang Medical College, Hei Long Jiang, China

3 Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden

4 Division of Orthopedics, Department for Clinical science, Intervention and technology (CLINTEC), Karolinska Institutet, Stockholm, Sweden

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Journal of Biomedical Science 2011, 18:59 doi:10.1186/1423-0127-18-59

Published: 19 August 2011

Abstract

Background

Developing efficient methods to isolate and identify human adipose-derived mesenchymal stem cells (hADSCs) remains to be one of the major challenges in tissue engineering.

Methods

We demonstrate here a method by isolating hADSCs from abdominal subcutaneous adipose tissue harvested during caesarian section. The hADSCs were isolated from human adipose tissue by collagenase digestion and adherence to flasks.

Results

The yield reached around 1 × 106 hADSCs per gram adipose tissue. The following comprehensive identification and characterization illustrated pronounced features of mesenchymal stem cells (MSCs). The fibroblast-like hADSCs exhibited typical ultrastructure details for vigorous cell activities. Karyotype mapping showed normal human chromosome. With unique immunophenotypes they were positive for CD29, CD44, CD73, CD105 and CD166, but negative for CD31, CD34, CD45 and HLA-DR. The growth curve and cell cycle analysis revealed high capability for self-renewal and proliferation. Moreover, these cells could be functionally induced into adipocytes, osteoblasts, and endothelial cells in the presence of appropriate conditioned media.

Conclusion

The data presented here suggest that we have developed high efficient isolation and cultivation methods with a systematic strategy for identification and characterization of hADSCs. These techniques will be able to provide safe and stable seeding cells for research and clinical application.