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多聚赖氨酸能增加用于细胞治疗的间充质干细胞产量

来源:生物谷 2016-07-14 20:21

2016年7月14日讯 /生物谷BIOON/ --间充质干细胞(MSC)是干细胞的一个研究分支,最初在骨髓中发现,因其具有多向分化潜能、造血支持和促进干细胞植入、免疫调控和自我复制等特点而日益受到人们的关注。

MSC拥有巨大的治疗潜力。MSC的临床研究已经在许多国家开展,美国批准了60余项临床试验,随着MSC及其相关技术的日益成熟,我国也批准了多项临床试验,走入了MSC核心技术研发的舞台,但是MSC在体外条件下的有效扩增是其临床应用的必须条件。众所周知胞外基质能够为其所包绕的细胞提供结构性和生化支撑,已经作为包被底物用于细胞培养。在这项研究中,来自韩国的研究人员旨在利用多聚赖氨酸在体外培养过程中增强MSC的功能性和干性。

他们通过细胞周期分析、分化实验、beta-半乳糖苷酶染色和RT-PCR的方法对MSC细胞的功能性进行分析。他们还进一步衡量了在无多聚赖氨酸包被和有多聚赖氨酸包被的培养板上生长的MSC的整体基因表达情况。结果发现在多聚赖氨酸包被过的培养板上生长的MSC细胞生长速率加快,并且干性标记物基因的表达发生上调。除此之外,他们的骨分化能力增强,参与细胞粘附、细胞周期、干性、细胞分化和细胞增殖的许多基因都出现上调。研究人员还证明生长在未包被培养板上的MSC细胞其beta-半乳糖苷酶表达高于生长在多聚赖氨酸包被培养板上的MSC细胞。

该研究表明多聚赖氨酸能够为MSC细胞培养提供良好的微环境,能够逆转复制衰老。这种方法能够显著提高用于细胞治疗的MSC细胞的产量。(生物谷Bioon.com)

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DOI: 10.1155/2016/8196078 

Poly-L-lysine Prevents Senescence and Augments Growth in Culturing Mesenchymal Stem Cells Ex Vivo.

Heo JS1, Kim HO2, Song SY3, Lew DH3, Choi Y1, Kim S

Mesenchymal stem cells (MSCs) possess great therapeutic potential. Efficient in vitro expansion of MSCs is however necessary for their clinical application. The extracellular matrix (ECM) provides structural and biochemical support to the surrounding cells, and it has been used as a coating substrate for cell culture. In this study, we have aimed to improve the functionality and stemness of MSCs during culture using poly-L-lysine (PLL). Functionality of MSCs was analysed by cell cycle analysis, differentiation assay, β-galactosidase staining, and RT-PCR. Furthermore, we assessed the global gene expression profile of MSCs on uncoated and PLL-coated plates. MSCs on PLL-coated plates exhibited a faster growth rate with increased S-phase and upregulated expression of the stemness markers. In addition, their osteogenic differentiation potential was increased, and genes involved in cell adhesion, FGF-2 signalling, cell cycle, stemness, cell differentiation, and cell proliferation were upregulated, compared to that of the MSCs cultured on uncoated plates. We also confirmed that MSCs on uncoated plates expressed higher β-galactosidase than the MSCs on PLL-coated plates. We demonstrate that PLL provides favourable microenvironment for MSC culture by reversing the replicative senescence. This method will significantly contribute to effective preparation of MSCs for cellular therapy.

 

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2016(第八届)干细胞技术与临床转化论坛

会议时间:2016.09.23-2016.09.24     会议地点:北京

会议详情: http://www.bioon.com/z/2016stemcell/

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