Mesenchymal Stem CellsDerivation and culture optimization of human mesenchymal stem cells for prospective transplantation therapies

Mesenchymal stem cells (MSCs) have emerged as a great promise in the field of regenerative medicine. These cells have stable genome and are able to self-renew and differentiate to multiple lineage of the mesenchyme. Their potential uses as therapeutic agents are based on their role in tissue repair and immunomodulation. These cells have great potential in treating many diseases including the graft vs host disease, heart disease, COVID-19 lung injury, diabetes, stroke, injury to spinal cord and many more(Han et al. 2019). However, some challenges need to be addressed and resolved before that. These cells need to be propagated in vitro to be produced in clinical grade as they are present in small numbers in the bone marrow, umbilical cord, adipose tissue and other sources. In vitro culture and expansion of MSCs needs to be done in conditions devoid of any animal or serum products to ensure safety of the patients after transplantation. Xeno-free culture conditions have been developed as the demand for the clinical application of human MSCs (hMSCs) has increased. We have shown in our work the suitability of xeno-free and serum-free culture conditions in stabilizing and promoting human embryonic stem cells (Albalushi et al. 2018) and human induced pluripotent stem cells (Panula et al. 2019) culture and expansion in vitro. However, in regard to hMSCs, studies are inconsistent about the influence of the xeno free culture conditions on the hMSCs. Studies demonstrated variation in the influence of xeno free culture conditions on hMSCs and recommended testing xeno free conditions prior to use for clinical application(Bui, Nguyen, and Than 2020). Moreover, extracellular matrix components have been shown to enhance and stabilize the characteristics of stem cells cultured in vitro. Among these component is laminin 521 which we have shown its positive effects on stabilizing and improving pluripotent stem cell culture (Albalushi et al. 2018).. Moreover, it has been used in culturing and differentiating pluripotent stem cells into many specific cells used now in clinical trials. In this study we aim to produce, for the first time in Oman, hMSCs from different resources in a condition permits their future use in the clinic. Human MSCs will be isolated and cultured in a culture condition devoid of any animal and serum products Furthermore, we aim to investigate the effect of laminin 521 as an initial step prior to shifting to the bioreactor for expansion of the cells in a GMP (Good Manufacturing practice) set up in the future. This study is expected to be completed in three years. It will establish the MSCs service in Oman, which can improve interventions such as management of COVID 19 and acute graft versus host disease.