One size does not fit all: developing a cell-specific niche for in vitro study of cell behavior.
One size does not fit all: developing a cell-specific niche for in vitro study of cell behavior.
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DOI：
10.1016/j.matbio.2016.01.004
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发表时间：
2016-05
期刊：
影响因子：
--
通讯作者：
Chen XD
中科院分区：
文献类型：
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作者：
Marinkovic M;Block TJ;Rakian R;Li Q;Wang E;Reilly MA;Dean DD;Chen XD
100多年来，细胞和组织一直在使用玻璃和塑料表面进行体外研究。在过去的10-20年里，大量的研究表明，细胞对它们的局部环境（细胞外基质，ECM）非常敏感，其中包含影响细胞行为的化学和物理线索。这些观察结果表明，使用组织培养聚苯乙烯（TCP）表面的现代细胞培养系统可能无法在体外重现真实的细胞行为，从而导致“人为结果”。在目前的研究中，我们使用骨髓（BM）和脂肪（AD）来源的基质细胞来制备BM-ECM和AD-ECM，它们在细胞合成后脱细胞，以模拟这些组织中每一种的细胞生态位。每种ECM的特征在于其影响BM-和AD-间充质干细胞（MSC）增殖以及三种癌细胞系（HeLa、MCF-7和MDA-MB-231）增殖、调节细胞扩散和相对于标准TCP表面直接分化的能力。我们发现，两种ECM都促进了MSC的增殖，但是当细胞的组织来源与ECM的组织来源相匹配时，这种效果得到增强（即BM-ECM促进BM-MSC的增殖超过AD-MSC，反之亦然）。此外，BM-和AD-ECM显示出分别优先引导MSC向成骨或成脂谱系分化，这表明ECM的作用是组织特异性的。此外，每种ECM影响细胞形态（即圆形），而与MSC的来源无关，这进一步支持了效应具有组织特异性的观点。有趣的是，与MSC不同，这些ECM不会促进癌细胞的增殖。为了进一步了解这三种培养基如何影响细胞行为，我们评估了两种ECM的化学（蛋白质组成）和物理特性（结构和机械）。虽然许多结构蛋白（如胶原蛋白和纤连蛋白）被发现在BM-和AD-ECM中的水平相当，架构（即纤维取向;表面粗糙度）和物理性质（储能模量，表面能）的每一个是独特的。这些结果表明，当在具有化学和物理性质差异的三种不同基质（BM-和AD-ECM和TCP）上培养时细胞行为的差异，提供了两种ECM可以概括骨髓和脂肪组织的天然干细胞生态位的特定元素的证据。更广泛地说，可以认为由细胞离体加工的ECM是开发组织特异性培养环境的理想起点。与TCP相比，TCP依赖于“一刀切”的范例，天然组织特异性ECM可能是一种更合理的模型，以接近工程化3D组织特异性培养系统来复制体内生态位。我们认为这种方法将为细胞行为的基础研究以及基于细胞的治疗提供更有意义的信息。
For more than 100 years, cells and tissues have been studied in vitro using glass and plastic surfaces. Over the last 10–20 years, a great body of research has shown that cells are acutely sensitive to their local environment (extracellular matrix, ECM) which contains both chemical and physical cues that influence cell behavior. These observations suggest that modern cell culture systems, using tissue culture polystyrene (TCP) surfaces, may fail to reproduce authentic cell behavior in vitro, resulting in “artificial outcomes.” In the current study, we use bone marrow (BM)- and adipose (AD)-derived stromal cells to prepare BM-ECM and AD-ECM, which are decellularized after synthesis by the cells, to mimic the cellular niche for each of these tissues. Each ECM was characterized for its ability to affect BM- and AD-mesenchymal stem cell (MSC) proliferation, as well as proliferation of three cancer cell lines (HeLa, MCF-7, and MDA-MB-231), modulate cell spreading, and direct differentiation relative to standard TCP surfaces. We found that both ECMs promoted the proliferation of MSCs, but that this effect was enhanced when the tissue-origin of the cells matched that of the ECM (i.e. BM-ECM promoted the proliferation of BM-MSCs over AD-MSCs, and vice versa). Moreover, BM- and AD-ECM were shown to preferentially direct MSC differentiation towards either osteogenic or adipogenic lineage, respectively, suggesting that the effects of the ECM were tissue-specific. Further, each ECM influenced cell morphology (i.e. circularity), irrespective of the origin of the MSCs, lending more support to the idea that effects were tissue specific. Interestingly, unlike MSCs, these ECMs did not promote the proliferation of the cancer cells. In an effort to further understand how these three culture substrates influence cell behavior, we evaluated the chemical (protein composition) and physical properties (architecture and mechanical) of the two ECMs. While many structural proteins (e.g. collagen and fibronectin) were found at equivalent levels in both BM- and AD-ECM, the architecture (i.e. fiber orientation; surface roughness) and physical properties (storage modulus, surface energy) of each were unique. These results, demonstrating differences in cell behavior when cultured on the three different substrates (BM- and AD-ECM and TCP) with differences in chemical and physical properties, provide evidence that the two ECMs may recapitulate specific elements of the native stem cell niche for bone marrow and adipose tissues. More broadly, it could be argued that ECMs, elaborated by cells ex vivo, serve as an ideal starting point for developing tissue-specific culture environments. In contrast to TCP, which relies on the “one size fits all” paradigm, native tissue-specific ECM may be a more rational model to approach engineering 3D tissue-specific culture systems to replicate the in vivo niche. We suggest that this approach will provide more meaningful information for basic research studies of cell behavior as well as cell-based therapeutics.