Cell sheet biofabrication by co-administration of mesenchymal stem cells secretome and vitamin C on thermoresponsive polymer
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Cell sheet technology aims at replacement of artificial extracellular matrix (ECM) or scaffolds, popular in tissue engineering, with natural cell derived ECM. Adipose tissue mesenchymal stem cells (ASCs) have the ability of ECM secretion and presented promising outcomes in clinical trials. As well, different studies found that secretome of ASCs could be suitable for triggering cell free regeneration induction. The aim of this study was to investigate the effect of using two bio-factors: secretome of ASCs (SE) and vitamin C (VC) for cell sheet engineering on a thermosensitive poly N-isopropyl acryl amide-Methacrylic acid (P(NIPAAm-MAA)) hydrogel. The results revealed that using thermosensitive P(NIPAAm-MAA) copolymer as matrix for cell sheet engineering lead to a rapid ON⁄OFF adhesion/deadhesion system by reducing temperature without enzymatic treatment (complete cell sheet release takes just 6 min). In addition, our study showed the potential of SE for inducing ASCs sheet formation. H&E staining exhibited the properties of a well-formed tissue layer with a dense ECM in sheets prepared by both SE and VC factors, as compared to those of VC or SE alone. Functional synergism of SE and VC exhibited statistically significant enhanced functionality regarding up-regulation of stemness genes expression, reduced β-galactosidase associated senescence, and facilitated sheet release. Additionally, alkaline phosphatase activity (ALP), mineralized deposits and osteoblast matrix around cells confirmed a better performance of ostogenic differentiation of ASCs induced by VC and SE. It was concluded that SE of ASCs and VC could be outstanding biofactors applicable for cell sheet technology.
This project was supported financially by Umbilical Cord Stem Cell Research Center, Tabriz University of Medical Science (Grant number: 5/104/1149).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 1.Alizadeh E, Akbarzadeh A, Eslaminejad MB, Barzegar A, Hashemzadeh S, Nejati‐Koshki K, et al. Up regulation of liver‐enriched transcription factors HNF4a and HNF6 and liver‐specific MicroRNA (miR‐122) by inhibition of Let‐7b in mesenchymal stem cells. Chem Biol Drug Des. 2015;85:268–79.CrossRefGoogle Scholar
- 4.Biazar E, Montazeri N, Pourshamsian K, Asadifard F, Ghorbanalinezhad E, Keshel SH. et al. Harvesting epithelial cell sheet based on thermo-sensitive hydrogel. J Paramed Sci. 2010;1:27–33.Google Scholar
- 5.Harimoto M, Yamato M, Hirose M, Takahashi C, Isoi Y, Kikuchi A, et al. Novel approach for achieving double‐layered cell sheets co‐culture: overlaying endothelial cell sheets onto monolayer hepatocytes utilizing temperature‐responsive culture dishes. J Biomed Mater Res. 2002;62:464–70.CrossRefGoogle Scholar
- 34.Klingemann H. Methods of use of culture supernatant obtained from mesenchymal stem cells from dogs and cats for treatment of organ dysfunction. Google Patents. 2014.Google Scholar
- 47.Altomare L, Cochis A, Carletta A, Rimondini L, Farè S. Thermo-responsive methylcellulose hydrogels as temporary substrate for cell sheet biofabrication. J Mater Sci: Mater Med. 2016;27:1–13.Google Scholar
- 68.Kondo M, Murakami D, Yamato M, Takagi R, Namiki H, Okano T. Serum-dependent epithelial cell sheet shrinkage upon detachment from temperature-responsive culture surfaces. FASEB J. 2009;23:468.5–.5.Google Scholar