Adipose-derived stem cells differentiate into osteoblasts and chondrocytes

doi:0.3969/j.issn.2095-4344.2013.14.008

Abstract

Abstract:

BACKGROUND: Adipose-derived stem cells can be separated and obtained from fat tissue. Fat tissue distributes in the whole body, and can be easily obtained in large quantities and has less damage to the donor site when drawing.
OBJECTIVE: To identify the methods of in vitro isolating and culturing of mice adipose-derived stem cells, to induce the adipose-derived stem cells to differentate into chondrocytes and osteoblasts and to investigate the feasibility of being seed cells in tissue engineering.
METHODS: The adipose-derived stem cells were isolated from the epididymal fat tissue of Kunming mice. Primary adipose-derived stem cells were obtained and purified by collagenase Ⅰ digestion and differential adherence method. The adipose-derived stem cells were induced with osteogenic induction medium, and then gomori alkaline phosphatase staining and alizarin red calcium nodules staining were performed to detect the differentiation of adipose-derived stem cells; the adipose-derived stem cells were induced with cartilage induction medium, and the toluidine blue staining, safranin-O staining and type Ⅱ collagen immunohistochemistry testing were performed to detect the differentiation of adipose-derived stem cells.
RESULTS AND CONCLUSION: The adipose-derived stem cells were spindle-shaped and in adherent growth. After primary cultured for 7-9 days, the cells could reach 90% confluence. After passaged to the third generation, the cell morphology was in consistency, and the growth curve of the passaged adipose-derived stem cells presented “S” shape. The expressions of CD29 and CD44 antigens were positive detected with cell-specific antigen test, but the expressions of CD34 and CD45 were negative. After osteoblast-inducing culture, the differentiation of adipose-derived stem cells towards osteoblasts was verified positively by alkaline phosphatase staining and alizarin red staining. After chondrocyte-inducing culture, the differentiation of adipose-derived stem cells into the chondrocyte was verified positively by oil red O staining, type Ⅱ collagen immunohistochemical staining and toluidine blue staining. The adipose-derived stem cells with differentiation potential can be isolated from the fat tissues, and can be the cells can be stably passaged and differentiated in vitro. The adipose-derived stem cells can be differentiated into osteoblasts and chondrocytes after induction, and the adipose-derived stem cells have the advantages of rich sources and easily obtained which can be considered as the ideal seed cells of tissue engineering.

Key words: stem cells, adipose-derived stem cells, cell culture, osteoblasts, chondrocytes, differentiation, alkaline phosphatase staining, alizarin red staining, type Ⅱ collagen, safranin-O staining, stem cell photographs-containing paper

CLC Number:

R394.2

Liu Xiao-tan, Xu Hai-bin, Lu Tan. Adipose-derived stem cells differentiate into osteoblasts and chondrocytes[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(14): 2525-2531.

Figures/Tables 5

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