Isolation, culture and identification of adipose-derived stem cells from mouse epididymis

doi:10.3969/j.issn.2095-4344.2014.28.018

Abstract

Abstract:

BACKGROUND: As a new kind of adult stem cells, adipose-derived stem cells get more and more attention, because of rich source, drawing materials easily and powerful proliferation.
OBJECTIVE: To isolate and culture adipose-derived stem cells from the epididymal adipose tissue in mice, and to identify their biological characteristics.
METHODS: Adipose tissue was obtained from epididymis in mice by aseptically cutting. The tissue was digested using collagenase. Adipose-derived stem cells were separated and purified by using one digestion, multiple collection method and differential adhesion method. The morphology of adipose-derived stem cells was observed using inverted microscopy and transmission electron microscopy. Growth curve of adipose-derived stem cells was drawn. Immunophenotype of adipose-derived stem cells was identified by flow cytometry. Adipose-derived stem cells were induced to differentiate into adipocytes and osteocytes using cell inductors. Compatibility of adipose-derived stem cells and collagen scaffold material was observed using scanning electron microscope.
RESULTS AND CONCLUSION: Adipose-derived stem cells exhibited long spindle-like or fibroblast-like appearance, grew intensively and arranged in scroll and fascicular shape. In vitro, adipose-derived stem cells could be passaged to passage 9 under the inverted microscope. Under the transmission electron microscope, adipose-derived stem cells showed abundant microvilli on the cell surface. The nuclei were big in size. Some organelles were seen in cytoplasma, such as mitochondria and rough endoplasmic reticulum. Adipose-derived stem cells expressed CD44 and CD29, did not express CD34. After inducing by inductor, many small lipid droplets were seen in the cytoplasm of adipose-derived stem cells. The small lipid droplets were dyed red with oil red O. After induction of osteogenic inductor, the boundary line among adipose-derived stem cells was not clear and the structure of cells was fuzzy in the growth-intensive areas. There were many strong refractive granular material deposits at that field after dyeing with alizarin red. Scanning electron microscope revealed that adipose-derived stem cells were spread on the collagen scaffold. Results suggested that adipose-derived stem cells isolated by this method could amplify in vitro and stably subcultured. Under a certain inducing condition, adipose-derived stem cells could differentiate into osteoblasts and adipocytes, which showed a good compatibility with collagen scaffold.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: stem cells, adipose tissue, epididymis, mice, inbred BALB C, cells, cultured

CLC Number:

R394.2

Zhang Jian-qing, Ji Jia-lin, Cui Xin-ming, Zhang Qi, Li Yan-ru. Isolation, culture and identification of adipose-derived stem cells from mouse epididymis [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(28): 4535-4541.

Figures/Tables 6

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