Zahara Mansoor
University of Colombo, Sri Lanka
Title: Differentiation of osteogenic cells from umbilical cord mesenchymal stem cells: comparing two enzyme digestion methods
Biography
Biography: Zahara Mansoor
Abstract
Mesenchymal Stem cells (MSCs) are plastic-adherent, fibroblast – like cells with specific surface phenotype, having ability to differentiate into osteoblasts, chondroblasts and adipocytes in-vitro. Umbilical cord (UC) is a readily available without ethical constraints, showing high proliferation rate and osteogenic potential. To derive MSCs from the human UC Wharton’s Jelly (WJ) and osteogenic differentiation was my main objective.Following obtaining ethical approval, five UCs from healthy mothers undergoing elective Caesarian sections were collected, cleaned with phosphate buffered saline, removed blood vessels,digested WJ in 0.5% collagenase 2-3 hours / 0.2% collagenase overnight and cultured in DMEM supplemented with 10% FBS, 1% L-glutamine and 1% penstrep at 37Ëš C in 5% CO2. Cells are passaged at 70%confluency. At fourth passage (P4), osteogenic differentiation medium was added following incubation .Culture maintained for 21 days and cells were stained with 2% Alizarin red and von kossastains.MSCs were determined and characterized using Trypan blue test, Flow cytometry, RT-PCR and karyotypic analysis.Cells were were positive for CD90, CD73 and CD105 and negative for CD34 and CD45 markers expressing Oct-4 and G6PD genes. Karyotypes depicted were normal. Alizarin red stain gave bright orange red and von kossa stain gave black-brown deposits demonstrating the presence of extracellular calcium deposits.UC-MSCS serves as a suitable source for osteogenic regeneration Gene expression demonstrated the embryonic origin of the MSCs which maintained genomic stability upto P4 stage.So my initatiative stem cell research in Sri Lanka improves the therapeutic potential in bone defects and opens up new perspectives for bone tissue engineering.