Native extracellular matrix, synthesized ex vivo by bone marrow or adipose stromal cells, faithfully directs mesenchymal stem cell differentiation.
Native extracellular matrix, synthesized ex vivo by bone marrow or adipose stromal cells, faithfully directs mesenchymal stem cell differentiation.
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DOI：
10.1016/j.mbplus.2020.100044
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发表时间：
2020-11
影响因子：
--
通讯作者：
Chen XD
中科院分区：
文献类型：
--
作者：
Marinkovic M;Tran ON;Block TJ;Rakian R;Gonzalez AO;Dean DD;Yeh CK;Chen XD
间充质干细胞(MSCs)对微环境(小生境)中的提示高度敏感，必须在体外概括这些提示才能研究其真实行为。在这项研究中，我们假设天然的骨髓(BM)和脂肪(AD)来源的细胞外基质(ECM)在控制MSC行为方面是独一无二的。为了验证这一点，我们比较了骨髓(BM)来源的MSCs在由BM产生的天然脱细胞ECM和AD基质细胞(即BM-和AD-ECM)上的增殖和分化。我们发现，这两种细胞外基质含有相似类型的胶原蛋白，但每种胶原蛋白的相对丰度不同。VI型胶原含量最高(≈占总胶原量的60%)，而I型胶原含量次之，≈为30%。这两种类型的胶原蛋白在两种ECM中的比例几乎相等。相反，XII型胶原在AD-ECM中几乎是唯一存在的，而IV型和V型仅在BM-ECM中存在。在物理和机械方面，BM-ECM比AD-ECM更粗糙、更僵硬，但粘附性较弱。在培养14d的 中，两种ECM对骨髓间充质干细胞增殖的支持作用均好于组织培养塑料，但MSC相关表面标志物在三种培养表面上的表达均相对较高。在BM-ECM上培养的BM-MSCs在成骨(OS)分化介质中的钙沉积显著增加，表明成骨，而在AD-ECM上培养的BM-MSCs在成脂(AP)分化介质中油红O染色显著增加，表明有脂肪生成。此外，BM-ECM培养显著增加BM-MSC对成骨诱导剂rhBMP-2的反应性，而AD-ECM培养则增强对成脂诱导剂罗格列酮的反应性。这些发现支持我们的假设，并表明BM-ECM和AD-ECM保留了其组织特异性微环境(NICE)特有的元素，这些元素促进了MSC在扩增过程中的分化状态(即“茎”)的保留，并直接对谱系特异性诱导剂的细胞反应。这项研究为精确控制MSC的命运提供了一种新的范例，为组织特异性细胞治疗提供了所需的细胞谱系。组织培养塑料不是研究细胞体外行为的合适表面。细胞来源的细胞外基质已被证明可以概括干细胞的微环境。这些ECM引导MSC对谱系特异性诱导介质和因子的反应。这种新的培养系统有助于在体外建立生理细胞行为的模型。
Mesenchymal stem cells (MSCs) are highly responsive to cues in the microenvironment (niche) that must be recapitulated ex vivo to study their authentic behavior. In this study, we hypothesized that native bone marrow (BM)- and adipose (AD)-derived extracellular matrices (ECM) were unique in their ability to control MSC behavior. To test this, we compared proliferation and differentiation of bone marrow (BM)-derived MSCs when maintained on native decellularized ECM produced by BM versus AD stromal cells (i.e. BM- versus AD-ECM). We found that both ECMs contained similar types of collagens but differed in the relative abundance of each. Type VI collagen was the most abundant (≈60% of the total collagen present), while type I was the next most abundant at ≈30%. These two types of collagen were found in nearly equal proportions in both ECMs. In contrast, type XII collagen was almost exclusively found in AD-ECM, while types IV and V were only found in BM-ECM. Physically and mechanically, BM-ECM was rougher and stiffer, but less adhesive, than AD-ECM. During 14 days in culture, both ECMs supported BM-MSC proliferation better than tissue culture plastic (TCP), although MSC-related surface marker expression remained relatively high on all three culture surfaces. BM-MSCs cultured in osteogenic (OS) differentiation media on BM-ECM displayed a significant increase in calcium deposition in the matrix, indicative of osteogenesis, while BM-MSCs cultured on AD-ECM in the presence of adipogenic (AP) differentiation media showed a significant increase in Oil Red O staining, indicative of adipogenesis. Further, culture on BM-ECM significantly increased BM-MSC-responsiveness to rhBMP-2 (an osteogenic inducer), while culture on AD-ECM enhanced responsiveness to rosiglitazone (an adipogenic inducer). These findings support our hypothesis and indicate that BM- and AD-ECMs retain unique elements, characteristic of their tissue-specific microenvironment (niche), which promote retention of MSC differentiation state (i.e. “stemness”) during expansion and direct cell response to lineage-specific inducers. This study provides a new paradigm for precisely controlling MSC fate to a desired cell lineage for tissue-specific cell-based therapies. Tissue culture plastic is an inadequate surface for studying cell behavior in vitro. Cell-derived ECMs have been shown to recapitulate the stem cell microenvironment. These ECMs direct MSC response to lineage-specific induction media and factors. This novel culture system facilitates modeling of physiologic cell behavior in vitro.