Three-dimensional spherical culture of adipose-derived stem cells

doi:10.3969/j.issn.2095-4344.2014.06.009

Abstract

Abstract:

BACKGROUND:Many types of mammalian cells aggregate and display three-dimensional multicellular spheroids when they are in normal physiological conditions. In order to observe and explore cellular natural states, many researchers try to use spherical cell culture in vitro, a common three-dimensional culture pattern.

OBJECTIVE:To use three different methods for spherical culture in vitro of adipose-derived stem cells and to observe their biological features.

METHODS: Adipose-derived stem cells were confirmed by the analysis of the markers for cell phenotypes as well as adipogenic and osteogenic differentiation potential assays. Three different methods of sphere cultures were used as follows: (1) ultra low attachment culture; (2) hanging-drop culture and (3) Eppendorf tube culture. The sphere formation was compared among above three methods. We used Imagej to calculate mean areas of these spheres. And we used Viability/Cytotoxicity Assay Kit for Animal Live & Dead Cells to detect their vitality.

RESULTS AND CONCLUSION: (1) Adipose-derived stem cells were confirmed by the analysis of the markers for cell phenotypes, CD29, CD44, CD59 were positive, as well as adipogenic and osteogenic differentiation potential assays were positive. The conventional monolayer cultures of adipose-derived stem cells showed spindle and cloning growth within three passages. (2) Ultra low attachment culture, hanging-drop culture, Eppendorf tube culture all could elicit adipose-derived stem cells spherical growth. However, spherical size, shape and uniformity differed depending on cell numbers, culture time and spherical culture methods. The ultra low attachment culture was comparatively difficult to control spherical shape and uniformity of adipose-derived stem cells. But hanging-drop culture and Eppendorf tube culture were able to form even cell spheres. (3) Spherical formation of adipose-derived stem cells using our three methods displayed good cell vitality.

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, cell culture techniques, spheroids, cellular

CLC Number:

R394.2

Wang Chan, Dai Ying, Guo Yong-long, Yang Yan, Liu Qing, Chen Jian-su. Three-dimensional spherical culture of adipose-derived stem cells[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(6): 872-879.

Figures/Tables 6

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