Derivation of mesenchymal stem cells from umbilical cord by comparing two enzyme digestion methods: Differentiation of osteogenic cells from UC MSCS

8^th International Conference on Tissue Science and Regenerative Medicine

September 11- 12, 2017 Singapore

Fathima Zahara Mansoor, H Goonasekara and V Dissanayake

University of Colombo, Sri Lanka

Posters & Accepted Abstracts: J Tissue Sci Eng

Abstract :

Aim: The aim of the study is to derive MSCs from the human UC Wharton�s Jelly (WJ) and osteogenic differentiation. Method: Umbilical cord (UC) is a readily available without ethical constraints, showing high proliferation rate and osteogenic potential. 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% Pen Strep at 37 oC 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 Kossa stains. MSCs were determined and characterized using Trypan blue test, flow cytometry, RT-PCR and karyotypic analysis. Findings: Cells 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. Conclusion & Significance: UC-MSCS serves as a suitable source for osteogenic regeneration. Gene expression demonstrated the embryonic origin of the MSCs which maintained genomic stability up to P4 stage. So this initiative stem cell research in Sri Lanka improves the therapeutic potential in bone defects and opens up new perspectives for bone tissue engineering.