Abstract
The effective long-term cryopreservation of human mesenchymal stem cells (MSCs) is an essential prerequisite step and represents a critical approach for their sustained supply in basic research, regenerative medicine, and tissue engineering applications. Therefore, attempts have been made in the present investigation to formulate a freezing solution consisting of a combination of Selaginella bryopteris water-soluble extract with and without dimethyl sulfoxide (Me2SO) for the efficient long-term storage of human umbilical cord blood- (hUCB-) derived MSCs. The cryopreservation experiment using the formulated freezing solution was further performed with hUCB MSCs in a controlled rate freezer. A significant increase in postthaw cell viability and cell attachment of MSCs was achieved with freezing medium containing Selaginella bryopteris water extract along with 10% Me2SO as compared to the freezing medium containing Me2SO (10% v/v) alone. Furthermore, the decreasing apoptotic events and reactive oxygen species production along with increasing expression of heat shock proteins also confirmed the beneficial effect of Selaginella bryopteris water extract. The beneficial effect of Selaginella bryopteris water extract was validated by its ability to render postpreservation high cell viability. In conclusion, the formulated freezing solution has been demonstrated to be effective for the standardization of cryopreservation protocol for hMSCs.
Highlights
In the last few decades, the advent of stem cell research has been a promising milestone in the field of cryobiology
The human umbilical cord blood- (hUCB-)derived mononuclear cells were initially cultured for isolation of mesenchymal stem cells (MSCs) utilizing their plastic adherence property
The conventional cryopreservation method using Me2SO is effective, cell viability and cell recovery of MSCs cryopreserved with Me2SO were always remained a critical issue due to Me2SO-induced cytotoxicity and neurodifferentiation in preserved MSCs [8, 9]
Summary
In the last few decades, the advent of stem cell research has been a promising milestone in the field of cryobiology. Mesenchymal stem cells (MSCs) are specialized cells with a wide range of multiple therapeutic properties like antiinflammatory, antiapoptotic, antifibrotic, and immunomodulatory along with tumor tropism and differentiation [1, 2]. These cells possess an extensive array of functional properties, and an account of which has been very well documented [3, 4]. As an alternative strategy to the current scenario of ineffective cryoprotectants, we propose to utilize the resurrection property of Selaginella bryopteris for the effective cryopreservation of MSCs
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