Regenerative Biomedicine Group, Blond McIndoe Research Laboratories, The University of Manchester, Manchester, United Kingdom.
Tissue Eng Part A. 2013 Feb;19(3-4):368-79. doi: 10.1089/ten.TEA.2012.0124. Epub 2012 Sep 24.
Since the first reports of induction of adipose-derived stem cells (ASC) into neuronal and glial cell phenotypes, expectations have increased regarding their use in tissue engineering applications for nerve repair. Cell adhesion to extracellular matrix (ECM) is a basic feature of survival, differentiation, and migration of Schwann cells (SC) during nerve regeneration, and fibronectin and laminin are two key molecules of this process. Interaction between ECM and SC-like differentiated ASC (dASC) could potentially improve the neurotrophic potential of the stem cells. We have investigated the effect of ECM molecules on SC-like dASC in terms of proliferation, adhesion, and cell viability. Fibronectin and laminin did not affect the proliferation of dASC when compared with cell adherent tissue culture plastic, but significantly improved viability and cell attachment when dASC were exposed to apoptotic conditions. To assess the influence of the ECM molecules on dASC neurotrophic activity, dASC were seeded onto ECM-coated culture inserts suspended above dorsal root ganglia (DRG) sensory neurons. Neurite outgrowth of DRG neurons was enhanced when dASC were seeded on fibronectin and laminin when compared with controls. When DRG neurons and dASC were in direct contact on the various surfaces there was significantly enhanced neurite outgrowth and coculture with laminin-conditioned dASC produced the longest neurites. Compared with primary SCs, dASC grown on laminin produced similar levels of neurite outgrowth in the culture insert experiments but neurite length was shorter in the direct contact groups. Anti β1 integrin blocking antibody could inhibit baseline and dASC evoked neurite elongation but had no effect on outgrowth mediated by laminin-conditioned dASC. ECM molecules had no effect on the levels of nerve growth factor and brain-derived neurotrophic factor secretion from dASC. The results of the study suggest that ECM molecules can significantly improve the potential of dASC for nerve regeneration.
自首次报道诱导脂肪来源干细胞（ASC）分化为神经元和神经胶质细胞表型以来，人们对其在神经修复组织工程应用中的应用寄予厚望。细胞黏附到细胞外基质（ECM）是施万细胞（SC）在神经再生过程中存活、分化和迁移的基本特征，纤连蛋白和层粘连蛋白是这一过程的两个关键分子。ECM 与类似于 SC 的分化 ASC（dASC）之间的相互作用可能会提高干细胞的神经营养潜能。我们研究了 ECM 分子对类似于 SC 的 dASC 的增殖、黏附和细胞活力的影响。与细胞黏附组织培养塑料相比，纤连蛋白和层粘连蛋白对 dASC 的增殖没有影响，但当 dASC 暴露于凋亡条件时，它们显著提高了细胞活力和黏附性。为了评估 ECM 分子对 dASC 神经营养活性的影响，将 dASC 接种到 ECM 包被的培养小室中，该小室悬挂在背根神经节（DRG）感觉神经元上方。与对照相比，当将 dASC 接种到纤连蛋白和层粘连蛋白上时，DRG 神经元的轴突生长得到增强。当 DRG 神经元和 dASC 直接接触各种表面时，轴突生长明显增强，与层粘连蛋白条件化的 dASC 共培养产生的轴突最长。与原代 SC 相比，在层粘连蛋白上生长的 dASC 在培养小室实验中产生类似水平的轴突生长，但在直接接触组中轴突长度较短。抗 β1 整合素阻断抗体可以抑制基线和 dASC 诱导的轴突伸长，但对层粘连蛋白条件化的 dASC 介导的生长没有影响。ECM 分子对 dASC 分泌神经生长因子和脑源性神经营养因子的水平没有影响。研究结果表明，ECM 分子可以显著提高 dASC 促进神经再生的潜力。
