Annals of Tropical Medicine and Public Health | 2021
Establishment and Immunological Characterization of Mesenchymal Stem Cells Derived from Human Umbilical Cord
Abstract
This study was designed to isolate mesenchymal stem cells (MSCs) from human umbilical cords (HUCs) by two methods. For this purpose, fresh Human Umbilical cords (n = 8) were obtained after cesarean section divided randomly in two groups each group have (n = 4). In enzymatic group, UCs was cut into 5 cm2 segments. The segments were cut longitudinally, and blood vessels were removed. The remaining pieces were digesting the tissue pieces with collagenase type I (0.2 mg/ml) for 3 h at room temperature with mild shaking. While, in Explants group, scrape the wharton’s jelly away from the blood vessels and inner epithelium of the subamnion using a scalpel, then remove the blood vessels to be sure collect any remaining perivascular then collected wharton’s jelly. Identification of MSCs achieved through their morphology, Immunophenotypic analysis and trypan blue dye exclusion test. MSCs have been isolated based on the ability of MSCs to adhere on plastic surfaces. MSCs reactivity to CD34 and CD105 were tested by immunocytochemistry. Isolated MSCs exhibited positive reactivity toward CD105 cell surface marker. MSCs were negative for the hematopoietic surface markers including CD34. The evaluation of living cells in different passages using trypan blue dye exclusion technique, the percentage of living cells obtained by two methods for different passages was estimated to be about 90%. However, the number of isolated cells was 1.5×10 6 and 1×10 6 cells in the Enzymatic and Explants groups, respectively.