Tissue Eng Regen Med.  2023 Dec;20(7):1133-1143. 10.1007/s13770-023-00568-3.

Enhancing Viability of Human Embryonic Stem Cells during Cryopreservation via RGD-REP-Mediated Activation of FAK/AKT/FoxO3a Signaling Pathway

• Kim JH
• Choi JI
• Che YH
• Sung SH
• Lee H
• Lee S
• Park JH
• Lee YI
• Lee YS
• Jeon WB
• Kim YJ

• KMID: 2548533
• DOI: http://doi.org/10.1007/s13770-023-00568-3

Abstract

BACKGROUND
Cryopreservation is a crucial method for long-term storage and stable allocation of human pluripotent stem cells (hPSCs), which are increasingly being used in various applications. However, preserving hPSCs in cryogenic conditions is challenging due to reduced recovery rates.
METHODS
To address this issue, the Arginine-Glycine-Aspartate (RGD) motif was incorporated into a recombinant elastin-like peptide (REP). Human embryonic stem cells (hESCs) were treated with REP containing RGD motif (RGDREP) during suspension and cryopreservation, and the survival rate was analyzed. The underlying mechanisms were also investigated.
RESULTS
The addition of RGD-REP to the cryopreservation solution improved cell survival and pluripotency marker expression. The improvement was confirmed to be due to the activation of the FAK-AKT cascade by RGD-REP binding to hESC surface interin protein, and consequent inhibition of FoxO3a. The inactivation of FoxO3a reduced the expression of apoptosis-related genes, such as BIM, leading to increased survival of PSCs in a suspension state.
CONCLUSION
RGD-REP, as a ligand for integrin protein, improves the survival and maintenance of hPSCs during cryopreservation by activating survival signals via the RGD motif. These results have potential implications for improving the efficiency of stem cell usage in both research and therapeutic applications.

Keyword

Human embryonic stem cell; In vitro culture; Extracellular matrix; RGD motif; Cryopreservation