Comparison of the Efficacy of Mesenchymal Stem Cells Derived from Total Human Umbilical Cord and Wharton s Jelly Explants

Background and Aim: An increased demand for MSCs in regenerativemedicine has made scientists prioritize the development of MSCisolation methods. The umbilical cord (UC) has become one of the majorsources of mesenchymal stem cells (MSCs). Several techniques have been devised for the dissociation of tissues for a primary culture that canaffect the quantity and quality of the isolated cells. Wharton’s jelly is oneof the most promising tissue sources for mesenchymal stem cells, but notthe only source. The aim of our study was to develop one of the mostappropriate methods for the isolation of mesenchymal stem cells fromthe entire umbilical cord.Methods: UC was divided into 10-cm pieces. After isolation vessels,both ends of each vessel were closed, to form a loop. The loops weremildly digested by collagenase type I, Enzyme activity was neutralized.Then the Wharton jelly was further cut into tiny pieces and digestedwith collagenase type I for 16 h. After mild digestion, vessel loops wereplaced in T75 flasks containing DMEM-LG. After partial digestion, thedigested Wharton jelly was centrifuged, after removing the supernatant,the tiny partially digested tissues were transferred into the same T75flasks containing the vascular loops. The UC pieces and the vascularloops were cultured in DMEM-LG. On the other hand, to compare themethods, MSCs were isolated from Wharton’s jelly by explanted methodtoo.Results: We could retrieve 6 to 10 million MSCs for 8 to 10 days ofprimary culture. Average time until desired cell confluency attained was9.2 days for our method, 20.7 days for explant method. The average ofharvested MSCs per 10-cm piece of UC was estimated to be 550,000and 150,000 in this method and explant method respectively. Also,Inverted microscopy revealed fibroblast-like morphology after 16 h forthis method and 72 h for the explant method. MSCs isolated by ourmethod express CD73, CD90, CD105, and CD44, but not CD34 andCD45 (hematopoietic markers) and CD31. The genes SOX2, OCT4, andNANOG are expressed in isolated MSCs. The capacity of these MSCs todifferentiate into adipocytes and osteocytes highlights their applicationin regenerative medicine.Conclusion: This method is simple, reproducible, and cost-efficient.Moreover, this method is suitable for the production of a large number ofhigh-quality MSCs from just one UC in less than a month, to be used forcell therapy in an 80-kg person.

Keywords:

Umbilical cord (UC) , Mesenchymal stem cells (MSCs) , Regenerative medicine