Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (40): 7509-7512.doi: 10.3969/j.issn.1673-8225.2010.40.022

Previous Articles     Next Articles

Ossification induction and phenotype of a special cell in culture process of rat adipose-derived stem cells

Wang Fu-ke1, Liu Liu1, He Xiao-guang1, Zhao De-ping2, Dai Xiao-ming1, Li Yi-song1   

  1. 1 Department of Head and Neck Surgery, 2 Experimental Center, First Affiliated Hospital, Kunming Medical College, Kunming  650032, Yunnan Province, China　
  • Online:2010-10-01 Published:2010-10-01
  • Contact: Liu Liu, Professor, Doctor, Doctoral supervisor, Department of Head and Neck Surgery, First Affiliated Hospital, Kunming Medical College, Kunming 650032, Yunnan Province, China liuliu3939@126.com
  • About author:Wang Fu-ke☆, Doctor, Lecturer, Department of Head and Neck Surgery, First Affiliated Hospital, Kunming Medical College, Kunming 650032, Yunnan Province, China wfk.04@126.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30960388*;the Special-Purpose Foundation Program of Department of Science and Technology of Yunnan Province, No. 2007C0007R*;the Doctor Innovation Foundation Program of Kunming Medical College, No. KM2007D03*

PDF

315

Abstract:

BACKGROUND: In previous experiment, a type of special cell that mixed fusiform adipose-derived stem cells (ADSCs) was found. A great quantity of polyhedral cells offer nest shape growth and take on cobblestone shape morphology. This is closely related cells to ADSCs, and it is necessary to clarify its nature.
OBJECTIVE: To determine the nature of the special cell by analysis of this specific phenotype and the induction of bone formation of these cells.
METHODS: Rat ADSCs were isolate and cultured by collagenaseⅠ, cultured by the medium containing fetal bovine serum. The immunofluorescence method was used to observe expressions and distributions of some special surface marker of the polygonal and fibroblastic cells, such as CD34, CD90, CD133 and Flk-1. Cells at passage 2 were obtained and induced by osteoinductive culture medium. Alkaline phosphatase (ALP) and Von Kossa staining were done on day 14 to test the characteristic of ossification.
RESULTS AND CONCLUSION:  ①Shape of cells cultured in fetal bovine serum (FBS) showed that a great quantity of polyhedral cells offered nest growth and took on cobblestone morphology, and the fusiform cells were visible. ②The fusiform cells were positive for CD90 and negative for CD34, CD133 and Flk-1; the polyhedral cells were positive for CD34 and Flk-1, and a part of polyhedral cells were positive for CD133, and negative for CD90. ③ In the process of ALP staining, a great quantity of polyhedral cells died, forming blank region; that of some fusiform cells was positive. ④The polyhedral cells died in the process of Von Kossa staining, presenting blank region; a part of fusiform cells formed red calcium nodus. The results have shown that the special cell is vascular endothelial cells in the culture process of rat ADSCs.

CLC Number: