Taki Tiraihi, Zeinab Najafi Gravari, Masoud Soleimani
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引用次数: 0
Abstract
Skin burn repair requires efficient regeneration in order to achieve proper healing. We applied a sequential culturing system 2D1w/3D2w protocol for differentiating human mesenvhymal stem cell derived from adipose tissue (hMSC-AT) into epithelial-like stem cells (ELSC). hMSC-AT were cultured, characterized by immunophenotyping, mesenchymal differentiation and induced in a 2D culture using epithelial differentiation medium containing ATRA, ITS, dexamethasone, EGF and FGF-2 for 7 days (2D1w), followed by seeding in a 3D culture using chitosan hydrogel combined with the same cocktail (3D2w) for 2 weeks. The immunostaining of the cells was done at different stages of induction in order to characterize the differentiated cells using pan cytokeratin and anti-cytokeratins 14 and 18 antibodies. The expression of cytokeratins 5, 10, 14, 18 and 19 was evaluated by RT-PCR. The toxicity of chitosan hydrogel and proliferation of ELSC in 3D chitosan hydrogel were evaluated by MTT test. The viability of ELSC in 3D culture was evaluated by acridine orange/propidium iodide staining. The hMSC-AT were immunopositive to CD90, CD105, CD73, CD45 and CD34. The ELSC in the 2D culture expressed cytokeratin 19 after 7 days, while the other cytokeratins were not expressed. At the second week (3D culture), all of the above markers were expressed except cytokerstin10, moreover, the viability and proliferation were 98.02 ± 1.34 and 5.45 ± 0.36%, respectively. The cytotoxicity assay demonstrates the biocompatibility of chitosan hydrogel. The study reveals that hMSC-AT, seeded on chitosan hydrogel, can be induced into ELSC in the presence of other inducers.
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The journal publishes papers on vast aspects of cell research, including morphology, biochemistry, biophysics, genetics, molecular biology, immunology. The journal accepts original experimental studies, theoretical articles suggesting novel principles and approaches, presentations of new hypotheses, reviews highlighting major developments in cell biology, discussions. The main objective of the journal is to provide a competent representation and integration of research made on cells (animal and plant cells, both in vivo and in cell culture) offering insight into the structure and functions of live cells as a whole. Characteristically, the journal publishes articles on biology of free-living and parasitic protists, which, unlike Metazoa, are eukaryotic organisms at the cellular level of organization.
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