* Tissue-Specific Extracellular Matrix Enhances Skeletal Muscle Precursor Cell Expansion and Differentiation for Potential Application in Cell Therapy.
* Tissue-Specific Extracellular Matrix Enhances Skeletal Muscle Precursor Cell Expansion and Differentiation for Potential Application in Cell Therapy.
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DOI：
10.1089/ten.tea.2016.0489
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发表时间：
2017-05
影响因子：
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通讯作者：
De‐ying Zhang;Yong Zhang;Yuanyuan Zhang;Hualin Yi;Zhan Wang;Rongpei Wu;D. He;G. Wei;Shicheng Wei;Yun Hu;Junhong Deng;T. Criswell;J. Yoo;Yu Zhou;A. Atala
中科院分区：
文献类型：
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作者：
De‐ying Zhang;Yong Zhang;Yuanyuan Zhang;Hualin Yi;Zhan Wang;Rongpei Wu;D. He;G. Wei;Shicheng Wei;Yun Hu;Junhong Deng;T. Criswell;J. Yoo;Yu Zhou;A. Atala
骨骼肌前体细胞（MPC）被认为是治疗由于损伤、疾病或年龄引起的骨骼肌功能障碍的细胞疗法的关键候选者。然而，由于在传统培养条件下这些细胞表型表达的变化，从小组织活检中体外扩增足够数量的功能性骨骼肌细胞具有挑战性。因此，本研究的目的是开发一种更好的培养系统，用于MPCs的扩增和肌分化，以进一步用于治疗。为此，我们开发了一种理想的组织脱细胞化方法，并比较了不同基质支持MPC生长和分化的能力。通过由蒸馏水、0.2 mg/mL DNA酶或5%胎牛血清组成的去细胞化方法产生猪源性骨骼肌和肝脏和肾脏细胞外基质（ECM）。将脱细胞基质进一步均质化、溶解并与肝素修饰的透明质酸基水凝胶（ECM-HA-HP）组合。当在单独的凝胶、ECM或每种ECM-HA-HP基质上生长时，评估人MPC的细胞增殖和肌源性分化能力。当在ECM-HA-HP基质上培养时，与测试的其他基质相比，人MPC增殖显著增强，其中在肌肉ECM-HA-HP（mECM-HA-HP）基质上的增殖最大。与其他凝胶-ECM底物相比，在mECM-HA-HP底物上分化的肌管的数量显著增加，以及表达特异性肌原性细胞标志物（即，肌球蛋白、结蛋白、myoD和myf 5）。总之，骨骼mECM-HA-HP作为培养基质可能由于其与体内环境的相似性而提供了最佳的培养微环境。这些数据表明，骨骼肌衍生的ECM凝胶用于人MPC的扩增和分化的潜在用途，用于骨骼肌功能障碍的基于细胞的治疗。
Skeletal muscle precursor cells (MPCs) are considered a key candidate for cell therapy in the treatment of skeletal muscle dysfunction due to injury, disease, or age. However, expansion of a sufficient number of functional skeletal muscle cells in vitro from a small tissue biopsy has been challenging due to changes in phenotypic expression of these cells under traditional culture conditions. Thus, the aim of the study was to develop a better culture system for the expansion and myo-differentiation of MPCs that could further be used for therapy. For this purpose, we developed an ideal method of tissue decellularization and compared the ability of different matrices to support MPC growth and differentiation. Porcine-derived skeletal muscle and liver and kidney extracellular matrix (ECM) were generated by decellularization methods consisting of distilled water, 0.2 mg/mL DNase, or 5% fetal bovine serum. Acellular matrices were further homogenized, dissolved, and combined with a hyaluronic acid-based hydrogel decorated with heparin (ECM-HA-HP). The cell proliferation and myogenic differentiation capacity of human MPCs were assessed when grown on gel alone, ECM, or each ECM-HA-HP substrate. Human MPC proliferation was significantly enhanced when cultured on the ECM-HA-HP substrates compared to the other substrates tested, with the greatest proliferation on the muscle ECM-HA-HP (mECM-HA-HP) substrate. The number of differentiated myotubes was significantly increased on the mECM-HA-HP substrate compared to the other gel-ECM substrates, as well as the numbers of MPCs expressing specific myogenic cell markers (i.e., myosin, desmin, myoD, and myf5). In conclusion, skeletal mECM-HA-HP as a culture substrate provided an optimal culture microenvironment potentially due to its similarity to the in vivo environment. These data suggest a potential use of skeletal muscle-derived ECM gel for the expansion and differentiation of human MPCs for cell-based therapy for skeletal muscle dysfunction.