Abstract

DETECTION OF BETA-ACTIN AND OSTEOCALCIN GENES EXPRESSION IN DIFFERENTIATED OSTEOBLASTS BY RT-PCR

Yaseen I. Mamoori, Zahraa K. Zedan* and Sally T. Younis

Abstract

This study was designed to evaluate osteogenic potential of bone marrow mesenchymal stem cells (BM-MSCs) isolated from mouse bone marrow in vitro. BM-MSCs were collected by flushing from bones and isolated based on its ability to adhere on plastic surfaces. Reactivity of BM- MSCs to CD105 and CD34 was tested by immunocytochemical assay. Isolated MSCs exhibited positive reactivity to CD105 and negative for the haematopoietic surface marker CD34. Differentiation of isolated MSCs into osteoblast was induced by osteogenic medium consisting of high glucose- DMEM supplemented with Ascorbic acid, dexamethasone and beta glycerophosphate disodium salt hydrate after 21 days. The presence of beta-actin and osteocalcin genes was confirmed in differentiated cells by Reverse transcriptase PCR (RT-PCR) on day 14.

Keywords: osteogenic, mesenchymal, immunocytochemical.