Isolation, Culture and characterisation of Adipose Tissue-Derived Mesenchymal Stem Cells (ADMSCs) by Explant-Enzymatic Methods

Aim: In this study, we use combination of enzymatic and explant methods, for isolation and culture of ADMSCs.
Adipose tissues dissected out from the abdominal region of male Wistar rats. Tissues were minced into small pieces (1-2 mm), and then treated with trypsin-EDTA 0.25% for 30 minutes at 37 ̊C in shaker-incubator. Enzymatic treated tissues were centrifuged and floating parts were cultured. Statistical analysis of the cells number was investigated using GraphPad Prismv6 software.
Results showed that ADMSCs isolated with our methods have growth characteristics similar to conventional methods. ADMSCs were maintained up to 10th passage and exhibited a homogeneous population in terms of appearance and morphology. We demonstrated that ADMSCs by these combination methods have mesenchymal characteristic markers (positive for CD90, CD44, CD73, CD105 and negative for CD35, CD45, CD11b).
In the current study, we showed that the combination of enzymatic and explant methods creates a simple, inexpensive and reproducible method for isolation and culture of ADMSCs.