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在热敏水凝胶中从眼睑脂肪来源干细胞诱导生成上皮样细胞。

Derivation of epithelial-like cells from eyelid fat-derived stem cells in thermosensitive hydrogel.

作者信息

Heidari Keshel Saeed, Rostampour Maryam, Khosropour Golbahar, Bandbon B Atefehsadat, Baradaran-Rafii Alireza, Biazar Esmaeil

机构信息

a Farabi Eye Hospital , Tehran University of Medical Sciences , Tehran , Iran.

b Department of Biomaterials Engineering , Tonekabon Branch, Islamic Azad University , Tonekabon , Iran.

出版信息

J Biomater Sci Polym Ed. 2016;27(4):339-50. doi: 10.1080/09205063.2015.1130406. Epub 2016 Jan 13.

Abstract

Injectable hydrogel is one of the great interests for tissue engineering and cell encapsulation. In the study, the thermosensitive chitosan/gelatin/β-glycerol phosphate (C/G/GP) disodium salt hydrogels were designed and investigated by different analyses. The eye fat-derived stem cells were used to evaluate the biocompatibility of hydrogels based on their phenotypic profile, viability, proliferation, and attachment ability. The results show that the sol/gel transition temperature of the C/G/GP hydrogel was in the range of 31.1-33.8 °C at neutral pH value, the gelation time was shortened, and the gel strength also improved at body temperature when compared with the C/GP hydrogel. In vitro cell culture experiments with eyelid fat-derived stem cells in hydrogel showed beneficial effects on the cell phenotypic morphology, proliferation, and differentiation. Microscopic figures showed that the eyelid fat stem cell were firmly anchored to the substrates and were able to retain a normal stem cell phenotype. Immunocytochemistry (ICC) and real-time-PCR results revealed change in the expression profile of eyelid fat stem cells grown with hydrogels when compared to those grown on control in epithelial induction condition. This study indicates that using chitosan/gelatin/β-glycerol phosphate hydrogel for cell culture is feasible and may apply in minimal invasive surgery in the future.

摘要

可注射水凝胶是组织工程和细胞封装领域备受关注的材料之一。在本研究中,通过不同分析方法设计并研究了热敏性壳聚糖/明胶/β-甘油磷酸(C/G/GP)二钠盐水凝胶。利用眼周脂肪来源的干细胞,基于其表型特征、活力、增殖和附着能力来评估水凝胶的生物相容性。结果表明,在中性pH值下,C/G/GP水凝胶的溶胶/凝胶转变温度在31.1-33.8°C范围内,与C/GP水凝胶相比,其凝胶化时间缩短,且在体温下凝胶强度也有所提高。在水凝胶中进行的眼周脂肪来源干细胞的体外细胞培养实验显示,对细胞表型形态、增殖和分化具有有益影响。显微镜图像显示,眼周脂肪干细胞牢固地锚定在基质上,并能够保持正常的干细胞表型。免疫细胞化学(ICC)和实时定量PCR结果显示,与在上皮诱导条件下在对照上生长的眼周脂肪干细胞相比,在水凝胶中生长的眼周脂肪干细胞的表达谱发生了变化。本研究表明,使用壳聚糖/明胶/β-甘油磷酸水凝胶进行细胞培养是可行的,未来可能应用于微创手术。

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