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在改性聚(L-丙交酯)箔上脂肪组织源性干细胞向血管平滑肌细胞的分化。

Differentiation of adipose tissue-derived stem cells towards vascular smooth muscle cells on modified poly(L-lactide) foils.

机构信息

Department of Biomaterials and Tissue Engineering, Institute of Physiology of the Czech Academy of Sciences, Videnska 1083, 142 20, Prague 4, Czech Republic.

Second Faculty of Medicine, Charles University, V Uvalu 84, 150 06, Prague 5, Czech Republic.

出版信息

Biomed Mater. 2021 Feb 18;16(2):025016. doi: 10.1088/1748-605X/abaf97.

Abstract

The aim of our research was to study the behaviour of adipose tissue-derived stem cells (ADSCs) and vascular smooth muscle cells (VSMCs) on variously modified poly(L-lactide) (PLLA) foils, namely on pristine PLLA, plasma-treated PLLA, PLLA grafted with polyethylene glycol (PEG), PLLA grafted with dextran (Dex), and the tissue culture polystyrene (PS) control. On these materials, the ADSCs were biochemically differentiated towards VSMCs by a medium supplemented with TGFβ1, BMP4 and ascorbic acid (i.e. differentiation medium). ADSCs cultured in a non-differentiation medium were used as a negative control. Mature VSMCs cultured in both types of medium were used as a positive control. The impact of the variously modified PLLA foils and/or differences in the composition of the medium were studied with reference to cell adhesion, growth and differentiation. We observed similar adhesion and growth of ADSCs on all PLLA samples when they were cultured in the non-differentiation medium. The differentiation medium supported the expression of specific early, mid-term and/or late markers of differentiation (i.e. type I collagen, αSMA, calponin, smoothelin, and smooth muscle myosin heavy chain) in ADSCs on all tested samples. Moreover, ADSCs cultured in the differentiation medium revealed significant differences in cell growth among the samples that were similar to the differences observed in the cultures of VSMCs. The round morphology of the VSMCs indicated worse adhesion to pristine PLLA, and this sample was also characterized by the lowest cell proliferation. Culturing VSMCs in the differentiation medium inhibited their metabolic activity and reduced the cell numbers. Both cell types formed the most stable monolayer on plasma-treated PLLA and on the PS control. The behaviour of ADSCs and VSMCs on the tested PLLA foils differed according to the specific cell type and culture conditions. The suitable biocompatibility of both cell types on the tested PLLA foils seems to be favourable for vascular tissue engineering purposes.

摘要

我们的研究目的是研究脂肪组织来源的干细胞(ADSCs)和血管平滑肌细胞(VSMCs)在各种改性聚(L-丙交酯)(PLLA)箔上的行为,即原始 PLLA、等离子体处理的 PLLA、接枝聚乙二醇(PEG)的 PLLA、接枝葡聚糖(Dex)的 PLLA 和组织培养聚苯乙烯(PS)对照。在这些材料上,通过在培养基中补充 TGFβ1、BMP4 和抗坏血酸(即分化培养基)将 ADSCs 生化分化为 VSMCs。在非分化培养基中培养的 ADSCs 用作阴性对照。在两种培养基中培养的成熟 VSMCs 用作阳性对照。研究了各种改性 PLLA 箔和/或培养基成分差异对细胞粘附、生长和分化的影响。当它们在非分化培养基中培养时,我们观察到 ADSCs 在所有 PLLA 样品上的粘附和生长相似。分化培养基支持在所有测试样品上的 ADSCs 表达特定的早期、中期和/或晚期分化标志物(即 I 型胶原、αSMA、钙调蛋白、 smoothelin 和平滑肌肌球蛋白重链)。此外,在分化培养基中培养的 ADSCs 在样品之间的细胞生长方面表现出显著差异,这与在 VSMCs 培养中观察到的差异相似。VSMCs 的圆形形态表明其对原始 PLLA 的粘附性较差,该样品的细胞增殖也最低。在分化培养基中培养 VSMCs 抑制了它们的代谢活性并减少了细胞数量。两种细胞类型在等离子体处理的 PLLA 和 PS 对照上形成最稳定的单层。ADSCs 和 VSMCs 在测试的 PLLA 箔上的行为根据特定的细胞类型和培养条件而不同。两种细胞类型在测试的 PLLA 箔上的合适生物相容性似乎有利于血管组织工程目的。

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