In vitro amplification and ultrastructure of adult adipose-derived stromal cells

doi:10.3969/j.issn.2095-4344.2013.10.003

Abstract

Abstract:

BACKGROUND: Adipose-derived stromal cells can be induced to differentiate into various cells in vitro. However, the best induced time of adipose-derived stromal cells has not been well understood.
OBJECTIVE: To speculate the best period for induced differentiation response through in vitro amplification curve and ultrastructure of adult adipose-derived stromal cells.
METHODS: Adipose tissue from liposuction surgery was digested and centrifuged. Cells were extracted, cultured and passaged in vitro. Morphological changes of cells were observed under the inverted phase contrast microscope; the growth curve of adipose-derived stromal cells was drawn by cell count method; immunohistochemistry and immunofluorescence staining were used to detect CD44, CD29 and CD34 expression; and the ultrastructure of adult adipose-derived stromal cells was observed under transmission electron microscope.
RESULTS AND CONCLUSION: Adipose-derived stromal cells grew rapidly. Cells displayed logarithmic growth at passage 3 after cultured for 5-7 days, and the number of which reached (5.32±0.03) ×10⁷/L after cultured for 8 days, then was stably kept, and the cells were in long-spindled shape. The positive rate of CD29 and CD44 was (84.35±9.73)% and (86.37±8.45)% respectively, whereas CD34 expression was negative by immunohistochemistry and immunofluorescence staining. Naive organelles were observed under the transmission electron microscope. As multiple stem cells, adipose-derived stromal cells, at passage 3 and 6 after cultured for 8-12 days, were most in number and stable in the shape and had the ultrastructures which enable cell differentiation and synthesize the required protein. This time is the best time for induced differentiation response.

Key words: stem cells, adipose-derived stem cells, adult adipose-derived stromal cells, in vitro culture, identification, ultrastructure, morphological characteristics, growth curve, CD44, CD29, stem cell photographs-containing paper

CLC Number:

R394.2

Lu Yan-hui, Yuan Xiao-dong, Ou Ya, Cai Ya-nan, Sun Qiao-yu. In vitro amplification and ultrastructure of adult adipose-derived stromal cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(10): 1724-1729.

Figures/Tables 4

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