Graduate Institute of Biomedical Materials and Tissue Engineering, School of Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei 11031, Taiwan.
International Master/Ph.D. Program in Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan.
Int J Mol Sci. 2020 Jul 9;21(14):4864. doi: 10.3390/ijms21144864.
The cell-derived extracellular matrix (ECM) is associated with a lower risk of pathogen transfer, and it possesses an ideal niche with growth factors and complex fibrillar proteins for cell attachment and growth. However, the cell-derived ECM is found to have poor biomechanical properties, and processing of cell-derived ECM into gels is scarcely studied. The gel provides platforms for three-dimensional cell culture, as well as injectable biomaterials, which could be delivered via a minimally invasive procedure. Thus, in this study, an adipose-derived stem cell (ADSC)-derived ECM gel was developed and cross-linked by genipin to address the aforementioned issue. The genipin cross-linked ADSC ECM gel was fabricated via several steps, including rabbit ADSC culture, cell sheets, decellularization, freeze-thawing, enzymatic digestion, neutralization of pH, and cross-linking. The physicochemical characteristics and cytocompatibility of the gel were evaluated. The results demonstrated that the genipin cross-linking could significantly enhance the mechanical properties of the ADSC ECM gel. Furthermore, the ADSC ECM was found to contain collagen, fibronectin, biglycan, and transforming growth factor (TGF)-β1, which could substantially maintain ADSC, skin, and ligament fibroblast cell proliferation. This cell-derived natural material could be suitable for future regenerative medicine and tissue engineering application.
细胞外基质（ECM）来源于细胞，与病原体转移的风险较低相关，并且具有理想的生态位，其中含有生长因子和复杂的纤维状蛋白，可促进细胞附着和生长。然而，细胞外基质的生物力学性能较差，且将细胞外基质加工成凝胶的研究甚少。凝胶为三维细胞培养以及可注射生物材料提供了平台，这些材料可通过微创程序进行输送。因此，在这项研究中，通过京尼平交联开发了一种脂肪来源干细胞（ADSC）衍生的 ECM 凝胶，以解决上述问题。通过包括兔 ADSC 培养、细胞片、脱细胞、冻融、酶消化、pH 中和以及交联在内的多个步骤来制备京尼平交联的 ADSC ECM 凝胶。评估了凝胶的理化特性和细胞相容性。结果表明，京尼平交联可显著增强 ADSC ECM 凝胶的机械性能。此外，ADSC ECM 中含有胶原蛋白、纤连蛋白、大软骨素蛋白聚糖和转化生长因子（TGF）-β1，可显著维持 ADSC、皮肤和韧带成纤维细胞的增殖。这种源自细胞的天然材料可能适用于未来的再生医学和组织工程应用。
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