Biological resources including proteins, cells, and tissues were confronted with both safe and stable preservation for practical use in biotechnological industry. Particularly, cell therapy for regenerative engineering is needed to restricted regulation and accurate preservation. Therefore, this study was investigated improved conditions of mesenchymal stem cells from human umbilical cord (hUCs) or aspirated adipose tissues (hATs) for clinical cell banks. Both cells were isolated according to standard operation procedure of Hurim BioCell Inc. and analyzed the inherent characteristics in passage 4. To compare the ability of experimental groups after cryopreservation, proliferation ability using calculated values and cytomorphological patterns of each experimental step were analyzed. Also proteins such as ice-binding protein or caspase inhibitor were applied to add the preservation medium of hUCs or hATs. Result of preservation solution with 20% serum was considered a positive group. Recovery rate and expansion results showed specific dosage and cell type-dependent differences in the experimental group. Chromosomal stability and multipotency of hUCs or hATs were expressed stable pattern after cryopreservation using advanced medium. As a result, these additives could be substituted for xenogenic sources in banking of hUCs or hATs.
Recently, human mesenchymal stem cells (MSCs) are attracting attention as a useful source for regenerative therapy. Controlled production of cell therapy requires the establishment and management of an accurate isolation, characterization and monitoring for quality assurance of developing MSCs mediated. In this study, we were confirmed maintenance of potency of isolated and cultured human umbilical cord (hUC)-MSCs during ex vivo expansion or after cryopreservation. Expression of their cell specific marker was analyzed by flow cytometry and the differentiation potency was confirmed by guided differentiation of adipocyte, osteocyte, chondrocyte and hepatocyte after expanding over 15 doublings in vitro. Safe production of developing a cell therapy was proved by testing for microbial, mycoplasma, endotoxin, and adventitious agents. Also stability of cells in cultivation, preservation and/or differentiation was determined chromosomal assay. In developing using hUC-MSCs, cells showed an accurate isolation and stable expansion in ex vivo condition. The results of several management assay showed that the stem cell marker expression of CD31, CD34 and CD45 were under 10%, however CD90 was over 90% by FACS analysis. Any contamination and mutation in all tests weren't detected in specific points for safe or stable production of hMC-MSCs. Also the proliferation and differentiation potency maintains during in vitro culture and after cryopreservation of hUC-MSCs. These results could be used as standard methods of maintenance of hUC-MSCs for cell therapy products and clinical application.