The study of ureteral decellularized matrix coating induced the differentiation of adipose-derived stem cells / 中华泌尿外科杂志

ABSTRACT

Objective:To investigate the effect of ureteral decellularized matrix (UDM) coating on the differentiation of adipose stem cells.Methods:From January 2018 to October 2019, UDM was prepared by perfusion method. H&E staining, DAPI staining, and DNA quantification were used to assessment the residues of cellular component in UDM and normal ureter; Masson′s trichrome staining and collagen quantification evaluated the collagen retention in UDM and normal ureter; the distribution and content of glycosaminoglycan in UDM and normal ureter were analyzed through Alcian Blue staining and glycosaminoglycan quantification. Canine adipose mesenchymal stem cells (cADMSCs) were isolated and cultured and identified by flow cytometry. The UDM was digested by pepsin enzyme to prepare the decellularized matrix coating as the experimental group. Type Ⅰ rat tail collagen coating was used as a control group. The cADMSCs were seeded on different coatings, and the differentiation of the cADMSCs was detected by immunofluorescence staining and Western Blot.Results:H&E and DAPI staining showed that the nuclear residue was not observed in the UDM. The DNA quantification demonstrated that the DNA content in UDM [(38.87±3.40) ng/mg] was significantly lower than that in the normal ureter group [(1 694.63±169.83) ng/mg, P<0.05]. Masson′s trichrome staining and collagen quantification confirmed that the collagen content in UDM [(265.89 ± 16.40) μg/mg] was no significantly different from the normal ureter group [(288.73 ± 16.32) μg/mg, P>0.05]. Alcian Blue staining showed the distribution of glycosaminoglycan in the UDM, and glycosaminoglycan quantification suggested that the content of glycosaminoglycan in the UDM [(1.57 ± 0.19) μg/mg] was lower than that in the normal ureter group [(3.43 ± 0.12) μg/mg] ( P<0.05). Immunofluorescence staining and Western Blot confirmed that the expression of Alpha-smooth muscle Actin (α-SMA) in the experimental group (2.51 ± 0.27, 3.68 ± 0.33, 4.91 ± 0.45) was higher than that in the control group (0.97±0.09, 1.02 ± 0.10, 1.00 ± 0.11) at 3 d, 7 d, 10 d ( P<0.05). The expression of α-SMA in the experimental group increased gradually with culture time ( P<0.05). While no changing of α-SMA expression in the control group was recordered. Conclusions:The prepared UDM removed the cellular components, and retained the collagen structure and bioactive components well; the ureter decellularized matrix coating could promote the differentiation of cADMSCs to smooth muscle cells.