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首页> 外文期刊>Acta biomaterialia >Covalent immobilization of stem cell factor and stromal derived factor 1α for in vitro culture of hematopoietic progenitor cells
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Covalent immobilization of stem cell factor and stromal derived factor 1α for in vitro culture of hematopoietic progenitor cells

机译：干细胞因子和基质衍生因子1α的共价固定化用于造血祖细胞的体外培养

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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.
机译：造血干细胞（HSC）目前用于血液疾病的治疗，但由于HSC的可用性有限，阻碍了HSC治疗剂的广泛应用。通过更好地了解HSC微环境以及精确概括其组件的能力，我们也许可以控制HSC行为。在这项工作中，我们开发了一种新型的仿生PEG水凝胶材料作为该目的的底物，并用不依赖于锚定的造血细胞系32D克隆3细胞测试了其潜力。我们固定了纤连蛋白衍生的粘附肽序列RGDS;对HSC自我更新至关重要的细胞因子，干细胞因子（SCF）;以及在HSC归巢和沉降中很重要的趋化因子，基质衍生因子1α（SDF1α）到聚（乙二醇）（PEG）水凝胶的表面。为了评估系统的功能，我们观察了生物分子对32D细胞粘附和形态的影响。我们证明了在表面上掺入RGDS可以剂量依赖性方式促进32D细胞粘附。我们还观察到与SCF或SDF1α结合的RGDS在表面附着力上的加和响应。此外，在含有固定的SCF或SDF1α的凝胶表面上，平均细胞面积增加而圆形度降低，表明细胞扩散增强。通过使用PEG水凝胶支架概括HSC微环境的各个方面，我们显示了控制32D细胞粘附和扩散的能力，并展示了该系统用于培养原代造血细胞群的潜力。

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