An efficient method for cell sheet bioengineering from rBMSCs on thermo-responsive PCL-PEG-PCL copolymer

被引:4
作者
Moghaddam, Sevil Vaghefi [1 ]
Abedi, Fatemeh [1 ]
Lotfi, Hajie [2 ]
Salehi, Roya [3 ]
Barzegar, Abolfazl [4 ]
Eslaminejad, Mohamadreza Baghaban [5 ]
Khalili, Mostafa [6 ]
Alizadeh, Effat [6 ]
机构
[1] Tabriz Univ Med Sci, Unit Tabriz Valiasr Hosp, Clin Res Dev, Tabriz, Iran
[2] Qazvin Univ Med Sci, Res Inst Prevent Noncommunicable Dis, Cellular & Mol Res Ctr, Qazvin, Iran
[3] Tabriz Univ Med Sci, Fac Adv Med Sci, Dept Med Nanotechnol, Tabriz, Iran
[4] Univ Tabriz, Fac Nat Sci, Dept Anim Biol, Tabriz, Iran
[5] ACECR, Royan Inst Stem Cell Biol & Technol, Cell Sci Res Ctr, Dept Stem Cells & Dev Biol, Tehran, Iran
[6] Tabriz Univ Med Sci, Fac Adv Med Sci, Dept Med Biotechnol, Tabriz, Iran
关键词
Cell sheets; PCL-PEG-PCL; Sol-Gel; Sodium Selenite; Stem cell; STROMAL CELLS; BLOCK-COPOLYMER; STEM-CELLS; HYDROGELS; SELENIUM; DIFFERENTIATION; FABRICATION;
D O I
10.1186/s13036-023-00346-8
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Utilizing both medium enrichment and a thermos-responsive substrate to maintain the cell-to-cell junctions and extracellular matrix (ECM) intact, cell sheet technology has emerged as a ground-breaking approach. Investigating the possibility of using sodium selenite (as medium supplementation) and PCL-PEG-PCL (as vessel coating substrate) in the formation of the sheets from rat bone marrow-derived mesenchymal stem cells (rBMSCs) was the main goal of the present study. To this end, first, Polycaprolactone-co-Poly (ethylene glycol)-co-Polycaprolactone triblock copolymer (PCEC) was prepared by ring-opening copolymerization method and characterized by FTIR, (1) H NMR, and GPC. The sol-gel-sol phase transition temperature of the PCEC aqueous solutions with various concentrations was either measured. Next, rBMSCs were cultured on the PCEC, and let be expanded in five different media containing vitamin C (50 mu g/ml), sodium selenite (0.1 mu M), vitamin C and sodium selenite (50 mu g/ml + 0.1 mu M), Trolox, and routine medium. The proliferation of the cells exposed to each material was evaluated. Produced cell sheets were harvested from the polymer surface by temperature reduction and phenotypically analyzed via an inverted microscope, hematoxylin and eosin (H&E) staining, and field emission scanning electron microscopy (FESEM). Through the molecular level, the expression of the stemness-related genes (Sox2, Oct-4, Nanog), selenium-dependent enzymes (TRX, GPX-1), and aging regulator gene (Sirt1) were measured by q RT-PCR. Senescence in cell sheets was checked by beta-galactosidase assay. The results declared the improved ability of the rBMSCs for osteogenesis and adipogenesis in the presence of antioxidants vitamin C, sodium selenite, and Trolox in growth media. The data indicated that in the presence of vitamin C and sodium selenite, the quality of the cell sheet was risen by reducing the number of senescent cells and high transcription of the stemness genes. Monolayers produced by sodium selenite was in higher-quality than the ones produced by vitamin C.
引用
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页数:15
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