Simple Production Method of Umbilical Cord Derived Mesenchymal Stem CelludUsing Xeno-Free Materials for Translational Research

摘要

Umbilical cord (UC) is a very promising source of mesenchymal stem cells (MSCs) for allogeneic use, as it can be collected easily without side effects to the donor. In this study, we developed a simple production protocol to get UC-MSCs using xeno-free material to provide safe MSCs for translational research in regenerative medicine. In this study, we used multiple harvest explant method to isolate the MSCs. The medium for isolation and propagation was 10% platelet concentrate (PC) containing alpha MEM. Confluent cultures were harvested, combined and counted. Primary cultures were expanded in T25 flasks (seeding around 5000/cm2) to passage-1 (P-1) and P-2, and the results of P-1 and P-2 cultures were counted. From 5 cm of umbilical cord, we did explant culture in four 24 well plates. The number of harvest per well ranges from 0-5 times i.e. no harvest= 6, once= 20, twice= 34, three times= 27, four times=8, and five times= 1 well(s), respectively. Therefore, we harvested a total of 206 times from the 96 wells, and got a total of 3,595,600 cells from the primary culture. If we use all of the cells from the primary culture for expansion, we will get a total of 47,809,462 cells in passage-1, a total of 1,022,607,122 cells in passage-2. In conclusion, UC-MSCs can be isolated and expanded easily in 10% PC containing alpha MEM, and is suitable to fill the demand of allogeneic MSCs for patient use.