Cultivation of adipose-derived mesenchymal stem cells from Rat Rattus norvegicus by using mechanical + enzymatic method

Abstract

Stem cells (stem cells) are the cells that do not have specific functions, can also differentiate into more than one type of cell that build tissues or organs of the body. In order to cultivate the stem cells from the primary tissue properly, it is necessary to optimize stem cells based on previous research methods. The design of this study was to cultivate mesenchymal stem cells from adipose (fat) tissue of Rat Rattus norvegicus by using mechanical + enzymatic method. The qualitative parameters that discussed were the option of handling isolation, time of incubation before cells reached confluence, and the morphology of mesenchymal stem cells obtained in each culture based on visual data using an inverted microscope. The quantitative parameters that analyzed were trypan blue assay and MTT assay. The results showed that the cultivation of mesenchymal stem cells were higher in the mechanical + enzymatic method compared with enzymatic methods. This method had a growth time from day 1 until it reached 80% confluence until day 7 and could be maintained until reached 90% confluence on day 9. Proliferation test by MTT assay showed the average of absorbance value was 0.942 with the cell number was 7.25 × 104 cells/µL. Further research is needed by flowcytometry assay and differentiation of this mesenchymal stem cells into osteocytes, adipocytes and chondrocytes.