Chondrogenic differentiation of human scalp adipose-derived stem cells in Polycaprolactone scaffold and using Freeze Thaw Freeze method

J Cell Physiol. 2018 Oct;233(10):6705-6713. doi: 10.1002/jcp.26477. Epub 2018 May 9.

Abstract

Human adipose tissue has been identified as a viable alternative source for mesenchymal stem cells. SADSCs were isolated from human scalp biopsy and then were characterized by Flow cytometry. SADSCS expressed CD90, CD44, and CD105 but did not express CD45 surface marker. Growth factors were used for chondrogenesis induction. Histology and immunohistology methods and gene expression by real-time PCR 14 days after induced cells have shown the feature of chondrocytes in their morphology and extracellular matrix in both inducing patterns of combination and cycling induction. Moreover, the expression of gene markers of chondrogenesis for example collagen type II aggrecan and SOX9 has shown by real-time PCR assay. Then, SADSCs were seeded alone on polycaprolatone (PCL) and with Freeze thaw Freeze (PCL+FTF) scaffolds and SADSCs differentiated toward the chondrogenic lineage and chondrogenesis induction were evaluated using scanning electron microcopy (SEM) and MTT assay. Our results showed that SADSCs were also similar to the other adipose-derived stem cells. Using TGF-beta3 and BMP-6 were effective for chondrogenesis induction. Therefore using of TGF-beta3 and BMP-6 growth factors may be the important key for in vitro chondrogenesis induction. The bio-composite of PCL+FTF nanofibrous scaffolds enhance the chondroblast differentiation and proliferation compared to PCL scaffolds .Therefore, our model will make it possible to study the mechanism of transition from chondroblast to chondrocyte.

Keywords: Freeze Thaw Freeze; Polycaprolactone; chondrogenic differentiation; scalp adipose-derived stem cells.

MeSH terms

  • Adipocytes / cytology
  • Adipocytes / drug effects
  • Adipose Tissue / cytology*
  • Adipose Tissue / drug effects
  • Adipose Tissue / growth & development
  • Bone Morphogenetic Protein 6 / genetics
  • Cell Differentiation / drug effects*
  • Cells, Cultured
  • Chondrocytes / cytology
  • Chondrocytes / drug effects
  • Chondrogenesis / drug effects
  • Extracellular Matrix / genetics
  • Humans
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects
  • Polyesters / pharmacology
  • Scalp / cytology*
  • Scalp / growth & development
  • Tissue Scaffolds
  • Transforming Growth Factor beta3 / genetics

Substances

  • Bone Morphogenetic Protein 6
  • Polyesters
  • Transforming Growth Factor beta3
  • polycaprolactone