Characterization of differentiation capacity of mesenchymal stem cells (MSCs) should be done before the MSCs can be used in regenerative medicine. Testing of differentiation capacity needs various substances containing medium, which are troublesome to prepare. Therefore, the objective of this study is to describe a simple and cheap method to check chondrogenic differentiation capacity of adipose tissue derived mesenchymal stem cells passage-5 by using fetal bovine serum (FBS) or platelet rich plasma (PRP) containing media in monolayer culture. In this study, adipose tissue derived mesenchymal stem cells (AT-MSCs) were isolated from lipoaspirate and cultured. To ensure the existence of AT-MSCs, flow cytometric analyses of passage-1 cells were done to count the CD34, CD73 and CD90 bearing cells. Prolonged cultures were done in various media, i.e. MesenCult®, 5% and 10% PRP containing high glucose Dulbeco's minimal Eagle's medium (HG-DMEM), 10% FBS containing α minimal essential medium (α MEM), and 10 ng/mL vascular endothelial growth factor (VEGF) and 10% human AB serum containing HG-DMEM. Ourf results showed that prolonged cultures in MesenCult® (one out of four), 5% PRP containing HG-DMEM (five out of six), 10% PRP containing HG-DMEM (all of nine), 10% FBS containing α MEM (twelve out of thirteen), and 10 ng/mL VEGF and 10% human AB serum containing HG-DMEM (seven out of eight) showed micromass formation. The micromasses were stained blue with alcian blue; therefore, micromass formation could be regarded as chondrogenic differentiation. In conclusion, prolonged culture in 5% PRP containing HG-DMEM, 10% PRP containing HG-DMEM, or 10% FBS containing α MEM can be used as an alternative cheap method to check the chondrogenic differentiation capacity of AT-MSCs.
|Number of pages||12|
|Journal||International Journal of PharmTech Research|
|Publication status||Published - 1 Jan 2014|
- Prolonged culture