Enhancement of keratinocyte growth factor potential in inducing adipose‐derived stem cells differentiation into keratinocytes by collagen‐targeting

Abstract

Different growth factors can regulate stem cell differentiation. We used keratinocyte growth factor (KGF) to direct adipose-derived stem cells (ASCs) differentiation into keratinocytes. To enhance KGF bioavailability, we targeted KGF for collagen by fusing it to collagen-binding domain from Vibrio mimicus metalloprotease (vibrioCBD-KGF). KGF and vibrioCBD-KGF were expressed in Escherichia coli and purified to homogeneity. Both proteins displayed comparable activities in stimulating proliferation of HEK-293 and MCF-7 cells. vibrioCBD-KGF demonstrated enhanced collagen-binding affinity in immunofluorescence and ELISA. KGF and vibrioCBD-KGF at different concentrations (2, 10, and 20 ng/ml) were applied for 21 days on ASCs cultured on collagen-coated plates. Keratinocyte differentiation was assessed based on morphological changes, the expression of keratinocyte markers (Keratin-10 and Involucrin), and stem cell markers (Collagen-I and Vimentin) by real-time PCR or immunofluorescence. Our results indicated that the expression of keratinocyte markers was substantially increased at all concentrations of vibrioCBD-KGF, while it was observed for KGF only at 20 ng/ml. Immunofluorescence staining approved this finding. Moreover, down-regulation of Collagen-I, an indicator of differentiation commitment, was more significant in samples treated with vibrioCBD-KGF. The present study showed that vibrioCBD-KGF is more potent in inducing the ASCs differentiation into keratinocytes compared to KGF. Our results have important implications for effective skin regeneration using collagen-based biomaterials.

3D Porous Polymer Scaffold-Conjugated KGF-Mimetic Peptide Promotes Functional Skin Regeneration in Chronic Diabetic Wounds

“…However, the KGF-mimetic peptide KGFp in our study did not alter the proliferative potential but significantly enhanced the migratory potential and differentiation of the MSCs toward keratinocytes. Like our findings, KGF-mediated differentiation of adipose-derived stem cells toward keratinocytes was demonstrated with an increased expression of the supra-basal and terminal keratinocyte genes, CK10 and Involucrin, respectively . ERK1/2 and AKT have been demonstrated as the major downstream effectors in KGF/FGFR2IIIB signaling , which correlates with our observation of activated of STAT3, ERK1/2, and AKT in regulating the transdifferentiation of MSCs toward KLCs.…”

Bioengineered MSCCxcr2 transdifferentiated keratinocyte-like cell-derived organoid potentiates skin regeneration through ERK1/2 and STAT3 signaling in diabetic wound

“…Keratinocytes express the receptor FGFR2IIIb, also known as Keratinocyte Growth Factor Receptor (KGFR) which binds to its cognate ligand FGF7 or KGF. The FGF7/FGFR2IIIb signaling regulates the proliferation, migration, and differentiation of skin keratinocytes [ 23 ]. The in silico analysis of the promoter sequence of mouse FGFR2IIIb ( KGFR ) revealed the presence of four putative proximal SP1 (p-ERK1/2) binding sites (−6 to −91) (Fig.…”