Evaluation of Sternness Maintenance Properties of the Recombinant Human Laminin α2 LG1-3 Domains in Human Mesenchymal Stern Cells

초록

Background: Laminin, a member of the Extracellular Matrix (ECM), is a glycoprotein that is used as a factor that affects cell adhesion, proliferation, survival, and differentiation. Of these, five globular domains (LG domains) of the alpha chain play an important role in influencing the cell by binding to the integrin. Objective: This study aimed to evaluate the ability of globular domains 1-3 of laminin alpha2 (rhLAMA2(LG1-3)) in maintaining the pluripotency of human Mesenchymal Stem Cells (hMSCs), which are widely used in regenerative medicine. Methods: hMSCs were grown in the medium supplemented with rhLAMA2(LG1-3), then the effect of the protein on hMSCs were confirmed through cell adhesion assay, proliferation assay and RT-PCR. Results: rhLAMA2(LG1-3) expressed in Escherichia coli has a molecular weight of 70 kDa, at 1 mu g/ml concentration of rhLAMA2(LG1-3), the attachment and proliferation of hMSCs were approximately 3.18-fold and 1.67-fold, respectively, more efficient than those of untreated controls. In addition, the undifferentiated state and degree of stemness of hMSCs were measured, on the basis of CD90 and CD105 levels. In the rhLAMA2(LG1-3)-treated hMSCs, the expression levels of CD90 and CD105 increased by 2.83-fold and 1.62-fold, respectively, compared to those in untreated controls. Conclusions: rhLAMA2(LG1-3) can be potentially used in stem cell therapy to improve the viability and maintain the undifferentiated state of hMSCs.

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