Connecting secretome to hematopoietic stem cell phenotype shifts in an engineered bone marrow niche
Connecting secretome to hematopoietic stem cell phenotype shifts in an engineered bone marrow niche
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DOI：
10.1093/intbio/zyaa013
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发表时间：
2020-07-01
影响因子：
2.5
通讯作者：
Harley, Brendan A. C.
中科院分区：
文献类型：
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作者：
Gilchrist, Aidan E.;Harley, Brendan A. C.
造血干细胞（HSC）主要存在于骨髓中，在那里它们从局部微环境中接收外部信号。生物物理信号、细胞成分和细胞分泌因子的复杂环境调节HSC产生血液和免疫系统的过程。我们先前表明，在甲基丙烯酰胺功能化明胶（GelMA）水凝胶中直接共培养原代小鼠造血干细胞和祖细胞与骨髓来源的间充质基质细胞和祖细胞（MSPC）群体可改善造血祖细胞的维持。然而，MSPCs影响HSC命运决定的机制仍然未知。在此，我们报告使用蛋白质组学分析，以关联HSC表型的一个广泛的候选池200可溶性因子产生的组合间充质和造血后代。偏最小二乘回归（PLSR），沿着迭代过滤方法，确定TGF β-1，MMP-3，c-RP和TROY与HSC维持呈正相关。在实验上，我们观察到用这些组合的细胞因子对GelMA水凝胶中的HSC单一培养物进行外源刺激，与未刺激的单一培养物相比，在7天培养后造血祖细胞与定向子代的比率增加7.52 +/-3.65倍。研究结果表明，向下选择的细胞因子的混合物放大了造血干细胞的造血维持潜力，超出了单独的MSPC分泌的因子。这项工作整合了经验和计算方法，以确定细胞因子的组合，以改善HSC的维护在一个工程HSC的小生境，提出了一个途径，确定无饲养层的培养平台，HSC的扩张。人工生态位内的造血干细胞以来自造血和间充质后代的可溶性因子的形式接受维持线索。应用蛋白质组学回归分析，我们确定了一组减少的可溶性因子相关的造血表型的维持在培养过程中的生物材料模型的骨髓龛。我们确定了一个最小的因素鸡尾酒，促进造血维持潜力的明胶为基础的文化，无论间充质饲养细胞的存在。通过结合经验和计算的方法，我们报告了一个实验可行的因素从一个大的数据集，使外源整合的可溶性因子到工程造血干细胞的静止干细胞群体的维持潜力增强。
Hematopoietic stem cells (HSCs) primarily reside in the bone marrow, where they receive external cues from their local microenvironment. The complex milieu of biophysical cues, cellular components and cell-secreted factors regulates the process by which HSC produce the blood and immune system. We previously showed direct coculture of primary murine hematopoietic stem and progenitor cells with a population of marrow-derived mesenchymal stromal and progenitor cells (MSPCs) in a methacrylamide-functionalized gelatin (GelMA) hydrogel improves hematopoietic progenitor maintenance. However, the mechanism by which MSPCs influenced HSC fate decisions remained unknown. Herein, we report the use of proteomic analysis to correlate HSC phenotype to a broad candidate pool of 200 soluble factors produced by combined mesenchymal and hematopoietic progeny. Partial least squares regression (PLSR), along with an iterative filter method, identified TGF beta-1, MMP-3, c-RP and TROY as positively correlated with HSC maintenance. Experimentally, we then observe exogenous stimulation of HSC monocultures in GelMA hydrogels with these combined cytokines increases the ratio of hematopoietic progenitors to committed progeny after a 7-day culture 7.52 +/- 3.65-fold compared to non-stimulated monocultures. Findings suggest a cocktail of the downselected cytokines amplifies hematopoietic maintenance potential of HSCs beyond that of MSPC-secreted factors alone. This work integrates empirical and computation methods to identify cytokine combinations to improve HSC maintenance within an engineered HSC niche, suggesting a route toward identifying feeder-free culture platforms for HSC expansion. Insight Hematopoietic stem cells within an artificial niche receive maintenance cues in the form of soluble factors from hematopoietic and mesenchymal progeny. Applying a proteomic regression analysis, we identify a reduced set of soluble factors correlated to maintenance of a hematopoietic phenotype during culture in a biomaterial model of the bone marrow niche. We identify a minimum factor cocktail that promotes hematopoietic maintenance potential in a gelatin-based culture, regardless of the presence of mesenchymal feeder cells. By combining empirical and computational methods, we report an experimentally feasible number of factors from a large dataset, enabling exogenous integration of soluble factors into an engineered hematopoietic stem cell for enhanced maintenance potential of a quiescent stem cell population.