微囊肿瘤检测：裸鼠胰腺癌模型评估。

Microencapsulated tumor assay: evaluation of the nude mouse model of pancreatic cancer.

机构信息

Department of General Surgery, Shanghai Institute of Digestive Surgery, Ruijin Hospital affiliated with to Shanghai JiaoTong University School of Medicine, Shanghai 200025, China.

出版信息

World J Gastroenterol. 2012 Jan 21;18(3):257-67. doi: 10.3748/wjg.v17.i3.257.

DOI:10.3748/wjg.v17.i3.257

PMID:22294829

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3261543/

Abstract

AIM

To establish a more stable and accurate nude mouse model of pancreatic cancer using cancer cell microencapsulation.

METHODS

The assay is based on microencapsulation technology, wherein human tumor cells are encapsulated in small microcapsules (approximately 420 μm in diameter) constructed of semipermeable membranes. We implemented two kinds of subcutaneous implantation models in nude mice using the injection of single tumor cells and encapsulated pancreatic tumor cells. The size of subcutaneously implanted tumors was observed on a weekly basis using two methods, and growth curves were generated from these data. The growth and metastasis of orthotopically injected single tumor cells and encapsulated pancreatic tumor cells were evaluated at four and eight weeks postimplantation by positron emission tomography-computed tomography scan and necropsy. The pancreatic tumor samples obtained from each method were then sent for pathological examination. We evaluated differences in the rates of tumor incidence and the presence of metastasis and variations in tumor volume and tumor weight in the cancer microcapsules vs single-cell suspensions.

RESULTS

Sequential in vitro observations of the microcapsules showed that the cancer cells in microcapsules proliferated well and formed spheroids at days 4 to 6. Further in vitro culture resulted in bursting of the membrane of the microcapsules and cells deviated outward and continued to grow in flasks. The optimum injection time was found to be 5 d after tumor encapsulation. In the subcutaneous implantation model, there were no significant differences in terms of tumor volume between the encapsulated pancreatic tumor cells and cells alone and rate of tumor incidence. There was a significant difference in the rate of successful implantation between the cancer cell microencapsulation group and the single tumor-cell suspension group (100% vs 71.43%, respectively, P = 0.0489) in the orthotropic implantation model. The former method displayed an obvious advantage in tumor mass (4th wk: 0.0461 ± 0.0399 vs 0.0313 ± 0.021, t = -0.81, P = 0.4379; 8th wk: 0.1284 ± 0.0284 vs 0.0943 ± 0.0571, t = -2.28, respectively, P = 0.0457) compared with the latter in the orthotopic implantation model.

CONCLUSION

Encapsulation of pancreatic tumor cells is a reliable method for establishing a pancreatic tumor animal model.

摘要

目的

利用癌细胞微囊化建立更稳定、更准确的胰腺癌裸鼠模型。

方法

本研究基于微囊化技术，即将人肿瘤细胞包裹在由半透膜构成的微小微囊（直径约 420μm）中。我们使用单个肿瘤细胞和包裹的胰腺肿瘤细胞注射，在裸鼠中建立了两种皮下植入模型。每周通过两种方法观察皮下植入肿瘤的大小，并根据这些数据生成生长曲线。在植入后 4 周和 8 周，通过正电子发射断层扫描-计算机断层扫描和尸检评估原位注射的单个肿瘤细胞和包裹的胰腺肿瘤细胞的生长和转移情况。然后将从每种方法获得的胰腺肿瘤样本送去进行病理检查。我们评估了癌症微囊与单细胞悬液相比，肿瘤发生率、转移存在率以及肿瘤体积和肿瘤重量的差异。

结果

体外连续观察微囊发现，微囊内的癌细胞在第 4 天至第 6 天增殖良好并形成球体。进一步的体外培养导致微囊膜破裂，细胞向外偏离并继续在培养瓶中生长。发现最佳注射时间是在肿瘤包裹后 5 天。在皮下植入模型中，包裹的胰腺肿瘤细胞与单独的细胞在肿瘤体积和肿瘤发生率方面均无显著差异。在同种异体植入模型中，癌细胞微囊化组与单个肿瘤细胞悬浮液组的成功植入率有显著差异（分别为 100%和 71.43%，P=0.0489）。在同种异体植入模型中，前者在肿瘤质量方面具有明显优势（第 4 周：0.0461±0.0399 比 0.0313±0.021，t=-0.81，P=0.4379；第 8 周：0.1284±0.0284 比 0.0943±0.0571，t=-2.28，P=0.0457）。