ObjectiveDifferentiation of immortalized Mesenchymal Stromal Cells (iMSCs) into PDGFR alpha-positive cells under controlled growth conditions has several vital implications in functional studies concerned with the pathogenesis of Diabetic Gastroparesis (DGP). A study published previously by our research group demonstrated the importance of these cells as a novel, in-vitro model for investigating the functional role of neuronal nitric oxide synthase. The currently available methods require fresh differentiation of PDGFR alpha-positive cells for each round of experimentation. This leads to longer delays, higher usage of reagents, and inconsistency in reproducibility of experiments frequently. We thus aimed to establish through validation that cryopreserving and maintaining the iMSC-derived PDGFR alpha-positive cells for functional investigations help us to overcome these challenges.ResultsWe demonstrated for the first time that the differentiated PDGFR alpha-positive cells from iMSCs can be cryopreserved and thawed to be used as per the experimental requirements with prolonged preservation of their characteristics. We assessed the viability of differentiated PDGFR alpha-positive cells pre- and post-freezing with the subsequent validation of their functional features using flow cytometry, qRT-PCR, and western blotting. We have been successful in demonstrating for the first time that the cryopreservation of previously differentiated PDGFR alpha-positive cells can be used as a feasible and cost-effective model for experimental reproducibility in functional studies of Diabetes Gastroparesis.
Funding Agencies|The authors would like to thank Mr. Manju N. J. (Sharjah Institute of Medical Research, University of Sharjah) for providing technical support in flow cytometry analysis.