Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (1): 29-32.doi: 10.3969/j.issn.1673-8225.2011. 01.006

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Isolation and culture characteristics and influential factors of Sprague Dawley rat adipose tissue-derived mesenchymal stem cells 

Hou Kai1, Li Mei1, Yang Yi-kun1, Li Chun1, Li Jin-ru1, Huang De-qing2   

  1. 1Basic Medical College, Tianjin Medical University, Tianjin  300070, China
    2Department of Orthopaedics, Tianjin Third Hospital, Tianjin  300250, China
  • Received:2010-09-01 Revised:2010-11-09 Online:2011-01-01 Published:2011-01-01
  • Contact: Huang De-qing, Doctor, Chief physician, Department of Orthopaedics, Tianjin Third Hospital, Tianjin 300250, China deqinghuang@yahoo.com.cn
  • About author:Hou Kai★, Studying for master’s degree, Basic Medical College, Tianjin Medical University, Tianjin 300070, China tjydhoukai@yahoo.com.cn
  • Supported by:

    the Application Basis and Advancing Research Project of Tianjin City in 2008, No. 08JCYBJC27100; the General Program of Science and Technology Foundation of Health Bureau of Tianjin City in 2007, No. 07KR01

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Abstract:

BACKGROUND: Adipose tissue-derived mesenchymal stem cells (AD-MSCs) could be obtained easily with abundant quantities; the fat cells left in the donation area have the capacity of proliferation and the injury is pretty mild. Therefore, AD-MSCs have been an ideal source of seed cells for tissue engineering. However, the isolation and culture of AD-MSCs still remain difficulties.
OBJECTIVE: To explore the method for isolation and culture of AD-MSCs and observe their growing properties.
METHODS: The adipose tissues were obtained from the inguinal groove, shoulder and abdominal cavity of Sprague Dawley rats aged 4 weeks and 10 months. Following washes and centrifugation, specimens were incubated in cell culture flask, and placed in 5%CO2 incubator. The liquid was replaced at 24, 48 and 72 hours. At a fusion of 80%, specimens were digested with 0.25% trypsogen-EDTA and passaged. Effects of age, gender, and the region of collecting adipose tissue, collagenase Ⅰ concentration and digestion time, and first time of liquid change on the culture of AD-MSCs were analyzed.
RESULTS AND CONCLUSION: Primary AD-MSCs presented spindle or multangular shape. While AD-MSCs passaged and cultured longer, the newborn AD-MSCs, with polygon shape in clusters, joined to pieces and stretched into spindle or multangular shape gradually. A few fat cells appeared after 48 hours in culture and the cell number increased when the culture time was prolonged. However, about 90% of the isolated cells still kept the shape of AD-MSCs. The AD-MSCs could be isolated successfully from the fat harvested from the 4-week female Sprague Dawley rat abdominal cavity, and be passaged stably in vitro. The Sprague Dawley rats’ age, sex, collection site of adipose tissue, the concentration of collagenase Ⅰ and its digestion time, as well as the time of the first change of medium have an impact on the isolation and culture of AD-MSCs.

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