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海藻酸钙包埋法微囊化软骨诱导分化骨髓间充质祖细胞用于细胞注射

Microencapsulation and chondrogenic differentiation of human mesenchymal progenitor cells from subchondral bone marrow in Ca-alginate for cell injection.

机构信息

TransTissue Technologies GmbH, Berlin, Germany.

出版信息

Acta Biomater. 2010 Feb;6(2):436-44. doi: 10.1016/j.actbio.2009.07.022. Epub 2009 Jul 19.

Abstract

The application of stem cells is a promising therapeutic approach for cartilage regeneration. For cell therapies, a biocompatible injectable carrier, which improves retention and cell distribution and enables cell differentiation, is a prerequisite. In this study, Ca-alginate microcapsules containing human subchondral cortico-spongious progenitor cells were prepared and the chondrogenic differentiation potential was verified by real-time reverse transcription-polymerase chain reaction analysis of typical chondrogenic marker genes. The results confirmed that these cells were able to differentiate along the chondrogenic lineage when encapsulated in Ca-alginate microcapsules with a mean diameter of 600-700microm and stimulated with TGF-beta3. Chondrogenic marker genes type II collagen, aggrecan and cartilage oligomeric matrix protein were induced together with type I collagen, whereas adipogenic and osteogenic marker genes showed no induction over 14 days. After 28 days, proteoglycans and type II collagen were evident histochemically and immunohistochemically. Mechanical stability as well as permeability of Ca-alginate capsules were analysed over the course of cultivation and found to be qualified for stable cell immobilization and sufficient exchange of solutes. Therefore, from the cell biology point of view, Ca-alginate, an established hydrogel scaffold material is suited for regenerative therapies of cartilage defects based on the injection of progenitor cells.

摘要

干细胞的应用是软骨再生有前途的治疗方法。对于细胞治疗,生物相容性可注射载体,它提高保留和细胞分布,并能使细胞分化,是一个前提。在这项研究中,含有人软骨下皮质-海绵状祖细胞的 Ca-藻酸盐微胶囊被制备,并通过实时逆转录聚合酶链反应分析典型的软骨形成标志物基因来验证其软骨分化潜能。结果证实,这些细胞能够在 TGF-β3 刺激下在直径为 600-700μm 的 Ca-藻酸盐微胶囊中沿着软骨谱系分化。软骨形成标志物基因 II 型胶原、聚集蛋白聚糖和软骨寡聚基质蛋白与 I 型胶原一起被诱导,而脂肪生成和成骨标志物基因在 14 天内没有被诱导。28 天后,在组织化学和免疫组织化学上可以明显看到蛋白聚糖和 II 型胶原。Ca-藻酸盐胶囊的机械稳定性和渗透性在培养过程中进行了分析,发现其适合于稳定的细胞固定和足够的溶质交换。因此,从细胞生物学的角度来看,Ca-藻酸盐,一种已建立的水凝胶支架材料,适合基于祖细胞注射的软骨缺陷的再生治疗。

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