Bioactive poly(ethylene glycol) hydrogels to recapitulate the HSC niche and facilitate HSC expansion in culture
Bioactive poly(ethylene glycol) hydrogels to recapitulate the HSC niche and facilitate HSC expansion in culture
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DOI：
10.1002/bit.25848
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发表时间：
2016-04-01
影响因子：
3.8
通讯作者：
West, Jennifer L.
中科院分区：
文献类型：
--
作者：
Cuchiara, Maude L.;Coskun, Sueleyman;West, Jennifer L.
造血干细胞（HSC）已经在治疗上使用了几十年，但是它们的广泛临床应用由于不能在体外成功地扩增HSC而受到阻碍。在培养中，HSC迅速分化并失去自我更新的能力。我们假设，通过模拟体外骨髓微环境的各个方面，我们可以更好地控制这些细胞的扩增和分化。在这项工作中，聚（乙二醇）二丙烯酸酯水凝胶的衍生物被用作造血干细胞和祖细胞（HSPC）群体的培养基质。关键的HSC细胞因子，干细胞因子（SCF）和干扰素（IFN），以及细胞粘附配体RGDS和连接段1共价固定到水凝胶的表面上。使用SCF和IFN，我们观察到HSPC的显著扩增，分别为约97和约104倍，同时保持c-kit（+）lin（-）和c-kit（+）Sca 1（+）lin（-）（KSL）群体以及在14天后形成多谱系集落的能力。HSPC也被封装在可降解的聚（乙二醇）水凝胶三维培养。在水凝胶中扩增后，约60%的细胞为c-kit（+），表明在14天的培养期内这些细胞的比例没有损失，形成的集落中约50%是多系的，表明细胞保留了其分化潜力。定制和使用该系统来支持HSC生长的能力可能会对HSC和其他血细胞类型在临床环境中的未来使用产生影响。Biotechnol. (c)2015 Wiley Periodicals，Inc.
Hematopoietic stem cells (HSCs) have been used therapeutically for decades, yet their widespread clinical use is hampered by the inability to expand HSCs successfully in vitro. In culture, HSCs rapidly differentiate and lose their ability to self-renew. We hypothesize that by mimicking aspects of the bone marrow microenvironment in vitro we can better control the expansion and differentiation of these cells. In this work, derivatives of poly(ethylene glycol) diacrylate hydrogels were used as a culture substrate for hematopoietic stem and progenitor cell (HSPC) populations. Key HSC cytokines, stem cell factor (SCF) and interferon- (IFN), as well as the cell adhesion ligands RGDS and connecting segment 1 were covalently immobilized onto the surface of the hydrogels. With the use of SCF and IFN, we observed significant expansion of HSPCs, approximate to 97 and approximate to 104 fold respectively, while maintaining c-kit(+)lin(-) and c-kit(+)Sca1(+)lin(-) (KSL) populations and the ability to form multilineage colonies after 14 days. HSPCs were also encapsulated within degradable poly(ethylene glycol) hydrogels for three-dimensional culture. After expansion in hydrogels, approximate to 60% of cells were c-kit(+), demonstrating no loss in the proportion of these cells over the 14 day culture period, and approximate to 50% of colonies formed were multilineage, indicating that the cells retained their differentiation potential. The ability to tailor and use this system to support HSC growth could have implications on the future use of HSCs and other blood cell types in a clinical setting. Biotechnol. (c) 2015 Wiley Periodicals, Inc.