Roberts Kevin, Kim John Taehwan, White Shelby, Hestekin Jamie, Wolchok Jeffrey C
Cell & Molecular Biology Program, University of Arkansas.
Department of Biomedical Engineering, University of Arkansas.
J Vis Exp. 2019 Feb 2(144). doi: 10.3791/58791.
Engineered scaffolds derived from extracellular matrix (ECM) have driven significant interest in medicine for their potential in expediting wound closure and healing. Extraction of extracellular matrix from fibrogenic cell cultures in vitro has potential for generation of ECM from human- and potentially patient-specific cell lines, minimizing the presence of xenogeneic epitopes which has hindered the clinical success of some existing ECM products. A significant challenge in in vitro production of ECM suitable for implantation is that ECM production by cell culture is typically of relatively low yield. In this work, protocols are described for the production of ECM by cells cultured within sacrificial hollow fiber membrane scaffolds. Hollow fiber membranes are cultured with fibroblast cell lines in a conventional cell medium and dissolved after cell culture to yield continuous threads of ECM. The resulting ECM fibers produced by this method can be decellularized and lyophilized, rendering it suitable for storage and implantation.
源自细胞外基质（ECM）的工程支架因其在加速伤口闭合和愈合方面的潜力而在医学领域引起了极大关注。体外从成纤维细胞培养物中提取细胞外基质有可能从人源以及潜在的患者特异性细胞系中生成细胞外基质，从而最大限度地减少异源表位的存在，而异源表位一直阻碍着一些现有细胞外基质产品的临床应用。体外生产适合植入的细胞外基质的一个重大挑战是，通过细胞培养生产细胞外基质的产量通常相对较低。在这项工作中，描述了在牺牲性中空纤维膜支架内培养细胞以生产细胞外基质的方案。将中空纤维膜与成纤维细胞系在常规细胞培养基中培养，细胞培养后溶解以产生连续的细胞外基质丝。通过这种方法产生的细胞外基质纤维可以进行脱细胞处理并冻干，使其适合储存和植入。
