Abstract
Objective To isolate and culture adipose-diverted stem cells (ADSCs) in SD rats using serial explant after enzymatic predigestion, and compare, the results with that from Zuk's method. Methods Adipose tissue from groin area of one-month-old SD rats was divided into two groups. In group A, the adipose tissue was snipped into 8mm3 blocks after washed with D-Hank's solution. The tissue blocks were digested by 0.25% trypsin for 5 minutes and 0.1% collagenase type 1 for 20 minutes, and put on 1mm grid nylon net for 3 to 4 days of cultivation. The tissues were then removed to another culture flask. They were repeatedly cultured by serial explant method for three or four times. In group B, the adipose tissue was treated by Zuk's method. MTT was used to measure bioactivity of the culture and flow cytometry was used to detect surface markers of the stem cells. Results In both groups, the cells had typical morphological characteristics of stem cells and expressed adipose stem cell surface markers CD105 and CD44. Conclusion The adipose-diverted stem cells cultured by technique of enzymatic digestion and serial explant in vitro have the same characteristics of those cultured with Zuk's method. This technique can yield plenty of ADSCs samples during a short period of time and thus enables its practical use in ADSCs cultivation. Key words: Stem cells; Cell culture techniques; Adipose; Explant method
Published Version
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