Although the implantation of intact tumor fragments is a common practice to generate orthotopic xenografts to study tumor invasion and metastasis, the direct implantation of tumor cell suspensions is necessary when prior manipulations of tumor cells are required. However, the establishment of orthotopic xenografts using tumor cell suspensions is not mature, and a comparative study directly comparing their engraftment and metastatic capabilities is lacking. It is unclear whether tumor fragments are superior to cell suspensions for successful engraftment and metastasis. In this study, we employed three GC cell lines with varying metastatic capacities to stably express firefly luciferase for monitoring tumor progression in real time. We successfully minimized the risk of cell leakage during the orthotopic injection of tumor cell suspensions without Corning Matrigel by systematically optimizing the surgical procedure, injection volume, and needle size options. Comparable high engraftment and metastatic rates between these two methods were demonstrated using MKN-45 cells with a strong metastatic ability. Importantly, our approach can adjust the rate of tumor progression flexibly and cuts the experimental timeline from 10–12 weeks (for tumor fragments) to 4–5 weeks. Collectively, we provided a highly reproducible procedure with a shortened experimental timeline and low cost for establishing orthotopic GC xenografts via the direct implantation of tumor cell suspensions.
尽管植入完整的肿瘤组织片段是生成原位异种移植瘤以研究肿瘤侵袭和转移的常用方法，但在需要对肿瘤细胞进行预先操作时，直接植入肿瘤细胞悬液则成为必要。然而，利用肿瘤细胞悬液建立原位异种移植瘤的技术尚不成熟，且缺乏直接比较其移植成功率和转移能力的对比研究。目前尚不清楚肿瘤组织片段是否在成功移植和转移方面优于细胞悬液。在本研究中，我们采用三种具有不同转移能力的胃癌细胞系，使其稳定表达萤火虫荧光素酶，以便实时监测肿瘤进展。通过系统优化手术步骤、注射体积和针头尺寸选择，我们在不使用康宁基质胶的情况下，成功将肿瘤细胞悬液原位注射过程中的细胞泄漏风险降至最低。利用具有强转移能力的MKN-45细胞，我们证明了这两种方法具有可比的高移植成功率和转移率。重要的是，我们的方法能够灵活调整肿瘤进展速率，并将实验周期从10-12周（针对肿瘤组织片段）缩短至4-5周。总之，我们提供了一种通过直接植入肿瘤细胞悬液建立胃癌原位异种移植瘤的高度可重复性方案，该方案具有缩短的实验周期和较低的成本。
肿瘤(癌症)患者之家