Covalent immobilization of stem cell factor and stromal derived factor 1α for in vitro culture of hematopoietic progenitor cells.
Covalent immobilization of stem cell factor and stromal derived factor 1α for in vitro culture of hematopoietic progenitor cells.
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DOI：
10.1016/j.actbio.2013.08.012
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发表时间：
2013-12
影响因子：
9.7
通讯作者：
West, Jennifer L.
中科院分区：
文献类型：
--
作者：
Cuchiara, Maude L.;Horter, Kelsey L.;Banda, Omar A.;West, Jennifer L.
造血干细胞(HSCs)目前被用于血液疾病的治疗，但由于HSCs的可获得性有限，HSC疗法的广泛应用受到了阻碍。通过更好地了解HSC的微环境以及准确概括其成分的能力，我们可能能够控制HSC的行为。在这项工作中，我们开发了一种新型的仿生聚乙二醇水凝胶材料作为底物，并用锚定非依赖的造血细胞系32D克隆3细胞测试了它的潜力。我们将纤维连接蛋白衍生的粘附肽序列RGDS、干细胞因子(SCF)和基质衍生因子1α(SDF1α)固定在聚乙二醇水凝胶的表面。为了评估该系统的性能，我们观察了生物分子对32D细胞黏附和形态的影响。我们证明了RGDS在表面的掺入以剂量依赖的方式促进了32D细胞的黏附。我们还观察到RGDS与干细胞因子或SDF1α联合使用时，表面粘附性的相加反应。此外，在含有固定化干细胞因子或SDF1α的凝胶表面，平均细胞面积增大，环形度减小，表明细胞铺展加快。通过使用聚乙二醇水凝胶支架重述HSC微环境的一些方面，我们已经展示了控制32D细胞的黏附和扩散的能力，并展示了该系统在原代造血细胞群体培养中的潜力。
Hematopoietic stem cells (HSCs) are currently utilized in the treatment of blood diseases, but widespread application of HSC therapeutics has been hindered by the limited availability of HSCs. With a better understanding of the HSC microenvironment and the ability to precisely recapitulate its components, we may be able to gain control of HSC behavior. In this work we developed a novel, biomimetic PEG hydrogel material as a substrate for this purpose and tested its potential with an anchorage independent hematopoietic cell line, 32D clone 3 cells. We immobilized a fibronectin-derived adhesive peptide sequence, RGDS; a cytokine critical in HSC self-renewal, stem cell factor (SCF); and a chemokine important in HSC homing and lodging, stromal derived factor 1α (SDF1α), onto the surfaces of poly(ethylene glycol) (PEG) hydrogels. To evaluate the system’s capabilities, we observed the effects of the biomolecules on 32D cell adhesion and morphology. We demonstrated that the incorporation of RGDS onto the surfaces promotes 32D cell adhesion in a dose dependent fashion. We also observed an additive response in adhesion on surfaces with RGDS in combination with either SCF or SDF1α. In addition, the average cell area increased and circularity decreased on gel surfaces containing immobilized SCF or SDF1α, indicating enhanced cell spreading. By recapitulating aspects of the HSC microenvironment using a PEG hydrogel scaffold, we have shown the ability to control the adhesion and spreading of the 32D cells and demonstrated the potential of the system for the culture of primary hematopoietic cell populations.
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影响因子：
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作者：
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通讯作者：
Smith, CW
影响因子：
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作者：
Cuchiara, Michael P.;Allen, Alicia C. B.;West, Jennifer L.
通讯作者：
West, Jennifer L.
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作者：
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通讯作者：
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作者：
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