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人脱落乳牙干细胞在三维丝素支架上的静态和动态培养分化研究。

A study of the differentiation of stem cells from human exfoliated deciduous teeth on 3D silk fibroin scaffolds using static and dynamic culture paradigms.

机构信息

Orthopedics Department, Mackay Memorial Hospital, Taipei, Taiwan.

PhytoHealth Corporation, Chin Sheng, Taiwan.

出版信息

Mater Sci Eng C Mater Biol Appl. 2020 Apr;109:110563. doi: 10.1016/j.msec.2019.110563. Epub 2019 Dec 16.

Abstract

Stem cells from human exfoliated deciduous teeth (SHED) are considered the best current source of human stem cells due to their ability to differentiate into multiple cell lineages. Dynamic co-culture systems can improve the culture environment, as they provide cells with signaling factors, extracellular matrixes, and cellular shear force, as well as enable the formation of heterotypic clusters. We seeded SHED in 3D silk fibroin porous scaffolds under static and dynamic cultures for 28 days, using the NIH3T3 cultivated medium as an induction agent. Many hepatospheres formed in these porous scaffolds, and cellular viability was shown to continually increase by MTT assays. Hepatic AFP and ALB gene expression, as well as glycogen storage, albumin secretion, and urea synthesis, were greater in cells in the 3D porous scaffold under a dynamic culture than in those cultured under 3D static culture and petri dish conditions. However, the 3D static culture is still superior to the traditional petri dish culture. The NIH3T3 cultivated medium can significantly induce hepatic differentiation of SHED, while the 3D dynamic culture system significantly enhances hepatic differentiation of SHED. This study provides alternative sources of hepatocytes for liver disease treatment.

摘要

人脱落乳牙来源的干细胞(SHED)因其能够分化为多种细胞谱系而被认为是目前人类干细胞的最佳来源。动态共培养系统可以改善培养环境,因为它们为细胞提供信号因子、细胞外基质和细胞剪切力,并促进异质簇的形成。我们在静态和动态培养条件下,将 SHED 接种在 3D 丝素多孔支架中 28 天,使用 NIH3T3 培养的培养基作为诱导剂。在这些多孔支架中形成了许多肝细胞球,通过 MTT 分析显示细胞活力持续增加。与 3D 静态培养和培养皿条件下培养的细胞相比,在 3D 动态培养下,多孔支架中的细胞的 AFP 和 ALB 基因表达以及糖原储存、白蛋白分泌和尿素合成更高。然而,3D 静态培养仍然优于传统的培养皿培养。NIH3T3 培养的培养基可以显著诱导 SHED 的肝分化,而 3D 动态培养系统可以显著增强 SHED 的肝分化。这项研究为肝脏疾病治疗提供了替代的肝细胞来源。

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