Okami Haruka, Muranushi Ryo, Yokobori Takehiko, Erkhem-Ochir Bilguun, Dorjkhorloo Gendensuren, Seki Takaomi, Okuyama Takayuki, Fukushima Ryousuke, Kawai Shunsuke, Hoshino Kouki, Dolgormaa Gantumur, Hagiwara Kei, Yamanaka Takahiro, Ishii Norihiro, Tsukagoshi Mariko, Igarashi Takamichi, Watanabe Akira, Kubo Norio, Araki Kenichiro, Saeki Hiroshi, Shirabe Ken
Department of General Surgical Science, Division of Hepatobiliary and Pancreatic Surgery, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511, Japan.
Division of Integrated Oncology Research, Gunma University Initiative for Advanced Research, Maebashi, Gunma 371-8511, Japan.
Exp Ther Med. 2024 Dec 24;29(2):39. doi: 10.3892/etm.2024.12789. eCollection 2025 Feb.
The present study aimed to investigate the role of a recombinant protein based on human collagen type I (RCPhC1) as a scaffold in maintaining the human tumor microenvironment within a patient-derived tumor xenograft (PDTX) model. RCPhC1, synthesized under animal component-free conditions, was explored for its potential to support the human-specific stroma associated with tumor growth. PDTX models were established using resected colorectal cancer liver metastasis specimens, and stromal cell populations from humans and mice were compared using three scaffolds: No scaffold (control), Matrigel and recombinant human collagen type I, across two passages. Specific antibodies for human Lamin B and mouse Lamin B were used for immunostaining to distinguish between human and mouse cells. Additionally, the impact of each scaffold on the invasive ability of mouse fibroblasts was assessed using an invasion assay. Patient-derived tumor tissues embedded with RCPhC1 hydrogels had significantly more human Lamin B-positive cells and fewer mouse Lamin B cells than those embedded with no scaffolds or Matrigel. The human Lamin B-positive cells in PDTX tumors with RCPhC1 hydrogels were recognized as fibroblasts. Additionally, these hydrogels significantly reduced the invasion of mouse fibroblast cell lines compared with Matrigel. The present study investigated RCPhC1 hydrogels as a new scaffold material for tumor engraftment in PDTX mouse models, and identified a promising experimental tool for maintaining the tumor microenvironment.
本研究旨在探讨基于人I型胶原蛋白的重组蛋白(RCPhC1)作为支架在患者来源的肿瘤异种移植(PDTX)模型中维持人肿瘤微环境的作用。在无动物成分条件下合成的RCPhC1,被研究其支持与肿瘤生长相关的人特异性基质的潜力。使用切除的结直肠癌肝转移标本建立PDTX模型,并在两个传代过程中使用三种支架:无支架(对照)、基质胶和重组人I型胶原蛋白,比较人和小鼠的基质细胞群体。使用针对人核纤层蛋白B和小鼠核纤层蛋白B的特异性抗体进行免疫染色,以区分人和小鼠细胞。此外,使用侵袭试验评估每种支架对小鼠成纤维细胞侵袭能力的影响。与无支架或基质胶包埋的患者来源肿瘤组织相比,包埋有RCPhC1水凝胶的患者来源肿瘤组织具有显著更多的人核纤层蛋白B阳性细胞和更少的小鼠核纤层蛋白B细胞。PDTX肿瘤中含RCPhC1水凝胶的人核纤层蛋白B阳性细胞被识别为成纤维细胞。此外,与基质胶相比,这些水凝胶显著降低了小鼠成纤维细胞系的侵袭。本研究将RCPhC1水凝胶作为PDTX小鼠模型中肿瘤移植的新型支架材料进行了研究,并确定了一种用于维持肿瘤微环境的有前景的实验工具。