β-galactosidase and Annexin V/PI Assays as QC Tests for Manufacturing cGMP-Grade Wharton Jelly Derived Mesenchymal Stem Cells

Abstract

Background: According to the guidelines for cGMP, strictly regulated procedures and quality control (QC) tests are required. However, none of the regulations by authorities suggested have β galactosidase and Annexin V/PI assays as QC tests. In this study, we aimed to demonstrate the feasibility of Annexin V and β galactosidase assays as valid cGMP compliant QC tests. Methods: Wharton’s Jelly derived mesenchymal stem cells (WJ MSCs) were isolated, characterized and screened as valid QC tests. In this study, in addition to routine QC tests, β gal assays were performed to confirm telomerase activity and Annexin V methods were used to confirm cell viability assays. Results: It was concluded that one donor (WJ MSC 3) has higher telomerase activity and it has lowest β gal activity at the same time (p<0,001). Only one donor showed significant difference in annexin V viability tests, indicating the importance of using automatic devices and Annexin V to manual tests for cell viability analysis (Muse: 92,48 ± 0,487% Flow Cytometry: 90,76 ± 1,116% p<0,01). Conclusion: With these existing data, telomerase activity and senescence levels may correlate and affect MSC quality in cell based therapy. In order to determine the viability values by adding Annexin V based automated methods could give more guaranteed results. Key words: Good Manufacturing Practice, SA b galactosidase, cell senescence, telomerase activity, Annexin V, cell based medicinal products. DOI: 10.7176/JSTR/6-12-07