Abstract:

BACKGROUND: Tissue culture technique was used to isolate human adipose-derived stem cells (ADSCs) for in vitro culture. This method is simple, can save time, and keeps biological properties of stem cells. According to this, the multiple-directional differentiation provides theoretic evidences and reference line for clinical application of tissue engineered seed cells. 
OBJECTIVE: To investigate the basic biological characteristics of human ADSCs by tissue culture techniques and their surface protein expression, and to explore their osteogenic and adipogenic potential in vitro.
METHODS: Human adipose tissue was obtained from the abdominal subcutaneous adipose tissue of health patients, and then ADSCs were isolated with tissue culture techniques and cultured in vitro. Morphology of ADSCs was observed. Cell cycle and surface proteins of the third passage of ADSCs were analyzed by flow cytometry technique. ADSCs from the third passages were induced into osteogenic and adipogenic lineages by different revulsants. Cells were examined by von Kossa staining and Oi1 Red O staining.
RESULTS AND CONCLUSION: Primary and passage ADSCs were fibroblast-like and could rapidly expand. The majority of the third passage of cells was in G0/G1 phase (88%). The third passage of ADSCs was positive for CD29, CD44, CD90 and CD105, while negative for CD34, CD45 and CD106. Human ADSCs were positive for Oi1 Red O staining at 21 days of adipogenic induction, and positive for Alizarin Reds staining and Von Kossa staining after osteogenic induction. Therefore, human ADSCs have the potential to differentiate into osteoblasts and adipocytes.