Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (10): 1764-1768.doi: 10.3969/j.issn.1673-8225.2011.10.012

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Influence of monoclonal screening on the purification of primary cultured rat adipose tissue-derived stem cells

Liu Jian1, 2, Li Li2, Ran Jiang-hua2, Zhang Sheng-ning2, Shao Jian-chun3   

  1. 1Depatrment of Surgery, Second Affiliated Hospital of Kunming Medical University, Kunming  650031, Yunnan Province, China; 2Department of Hepatobiliary and Pancreatic Surgery, Affiliated Ganmei Hospital of Kunming Medical University, Kunming  650011, Yunnan Province, China; 3Department of Laboratory Medicine, Affiliated Ganmei Hospital of Kunming Medical University, Kunming  650011, Yunnan Province, China
  • Received:2010-11-28 Revised:2011-01-18 Online:2011-03-05 Published:2011-03-05
  • Contact: Li Li, Professor, Chief physician, Department of Hepatobiliary and Pancreatic Surgery, Affiliated Ganmei Hospital of Kunming Medical University, Kunming 650011, Yunnan Province, China ynkmlili@yahoo.com
  • About author:Liu Jian☆, Studying for doctorate, Attending physician, Department of Surgery, Second Aftiliated Hospital, Kunming Medical University, Kunming 650031, Yunnan Province, China; Department of Hepatobiliary and Pancreatic Surgery, Affiliated Ganmei Hospital of Kunming Medical University, Kunming 650011, Yunnan Province, China Liujianwxc1219@163.com
  • Supported by:

    the Foundation of Science and Technology Department of Yunnan Province, No. 2007CA007*

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Abstract:

BACKGROUND: It was found that a kind of mesenchymal stem cell can be isolated from animal adult adipose tissues, that is, adipose tissue-derived stem cells (ADSCs). However, there were still some problems need to be improved of the traditional isolation methods.
OBJECTIVE: To investigate a new method that using monoclonal screening in the purification of primary cultured ADSCs. 
METHODS: After isolated the Lewis rat ADSCs with the digestion centrifuge process, the ADSCs were screened with the limited density dilution cloning and condition culture medium screening in the original generation culture. The group with clone screening ADSCs served as the experimental group, without clone screening as the control group. Cellular appearance, growth curve and cell doubling generation time were compared between groups. The surface markers CD49d, CD29, CD44 of ADSCs were determined by flow cytometry.
RESULTS AND CONCLUSION: The appearance of ADSCs which underwent clone screening had been homogeneously, with vigorous growing power and short doubling generation time. The growth curve was not changed obviously after many generations. The surface marks CD49d, CD29, and CD44 were expressed in both groups at 3-6 passages. The clone screening purification method is effective, economical purification method of ADSCs.

CLC Number: