此前，我们报道了使用三维（3D）工程细胞外基质（EECM）纤维支架结构成功地对小鼠结直肠癌细胞系（CT-26）进行细胞扩增。 CCL-247 在 3D EECM 或组织培养聚苯乙烯 (TCPS) 上在 8 天的有限时间内生长。然后分析细胞的生长、电穿孔效率和 Vigil 制造放行标准。使用 EECM 支架，我们报告了结直肠癌细胞系 CCL-247 (HCT116) 的扩增，从每个支架 2.45 × 105 个细胞的起始浓度扩展到 1.9 × 106 个细胞。扩展后，根据用于 FDA IIb 期试验的 Vigil 制造工艺的临床放行标准对 3D EECM 衍生细胞进行了评估。 3D EECM 衍生细胞通过了所有 Vigil 制造发布标准，包括细胞因子表达。在这里，我们展示了 Vigil 产品的成功制造，达到了 Vigil 治疗临床试验产品发布所需的规格。我们的结果证实，3D EECM 可用于扩增人类癌细胞 CCL-247，证明了涉及人体组织样本制造（包括芯针活检和最小腹水样本）的进一步临床开发的合理性。© 2024。作者。

Previously, we reported successful cellular expansion of a murine colorectal carcinoma cell line (CT-26) using a three-dimensional (3D) engineered extracellular matrix (EECM) fibrillar scaffold structure. CCL-247 were grown over a limited time period of 8 days on 3D EECM or tissue culture polystyrene (TCPS). Cells were then assayed for growth, electroporation efficiency and Vigil manufacturing release criteria. Using EECM scaffolds, we report an expansion of CCL-247 (HCT116), a colorectal carcinoma cell line, from a starting concentration of 2.45 × 105 cells to 1.9 × 106 cells per scaffold. Following expansion, 3D EECM-derived cells were assessed based on clinical release criteria of the Vigil manufacturing process utilized for Phase IIb trial operation with the FDA. 3D EECM-derived cells passed all Vigil manufacturing release criteria including cytokine expression. Here, we demonstrate successful Vigil product manufacture achieving the specifications necessary for the clinical trial product release of Vigil treatment. Our results confirm that 3D EECM can be utilized for the expansion of human cancer cell CCL-247, justifying further clinical development involving human tissue sample manufacturing including core needle biopsy and minimal ascites samples.© 2024. The Author(s).