Proliferation and differentiation of human mononucleated cells into osteoblast and osteoclast cells

Abstract

Stem cell is defined as the ability of the cell to proliferate themself and differentiate into more than one type of cells. Human mononucleated cells (MN cells) is a suspension cell that were isolated from peripheral blood that were originated from monocyte- machrophage lineage. The cells were cultured in complete media (CM) which consist of Alpha modified Essential Medium (AMEM) with 2% Penstrep and 10% Newborn calf serum (NBCS) for 30 days showed a significant increment (p<0.05) of viable cells compared to day 0 (n=3). The respective cells were differentiated at day 3 after in vitro proliferation into osteoblastic (OIB) and osteoclastic (OIC) cells by adding 10 nM /3- glycerophosphate and 50 ngluL ascorbic acid to induce osteoblast differentiation. In addition, human 50 ngluL sRANKL and 25 ng/ul. M-CSF were added to induce osteoclastic differentiation ofMN cells. Cells that were cultured in CM serve as a control and were subjected to the same approach as diferentiated cells. The increment of ALP enzyme for OIB differentiation and TRAP enzyme for OIC differentiation were evaluated via enzymology assay until day 14 and day 10 for OIB and OIC, respectively. ALP and TRAP enzyme showed a significant increase (p<O. (5) when differentiated for day 14 and JO compared to control cell, respectively. As a conclusion, human mononucleated cells is believe to have the potential to be defined as multi potent stem cells based on their fulfillment of a stem cell characteristic.