Method for building fluorescent tumor model in nude mice based on primary cells of human cerebral glioma

Abstract

The invention discloses a method for building a fluorescent tumor model in nude mice based on primary cells of human cerebral glioma, and belongs to the field of animal models in the medical fields. The method comprises the following steps: S1, culturing of steady-transferring cells: carrying out cell culturing by adopting an improved DMEM / F12 complete culturing medium, and massively amplifying the primary cells capable of stably expressing luciferase; S2, preparation of cell suspensions: repeatedly cleaning the cells by using the improved DMEM / F12 complete culturing medium, collecting the cell suspensions for subsequent use to ensure that the concentration of the cell suspensions is 6*10<9> cells / mL, and using the suspensions in 40 minutes after the suspensions are prepared; and S3, building of a glioma model of the nude mice: injecting the suspensions in craniums of the nude mice in situ to obtain the mice model capable of stably expressing the luciferase. The fluorescent tumor modelin nude mice based on the human cerebral glioma prepared by adopting the method has the advantages that tumor growth and pathological characteristics of the nude mice are similar to the human cerebral glioma, and growth of the tumors can be observed in real time, so that a desired research model is provided for researching glioma based on in-situ transplanting and tracing of the human cerebral glioma and researching micro-environments of the tumors.

Description

technical field [0001] The invention belongs to the field of animal models used in the medical field, and more specifically relates to a method for establishing a fluorescent nude mouse tumor model based on primary cells of human brain glioma. Background technique [0002] Glioma is a primary tumor derived from glial cells and their precursors. It is the most common and most difficult to treat malignant tumor of the central nervous system, accounting for about 50% of intracranial tumors. Due to the aggressive growth of most gliomas and the unclear boundary with the surrounding normal brain tissue, it is difficult to completely remove the tumor by surgery. Based on the fact that the biodiversity and pathogenesis of glioma cells have not yet been clarified, it is still a type of central nervous system tumor with the worst prognosis among all malignant tumors under comprehensive treatment. The average survival cycle of glioblastoma patients is only for 14.6 months. How to imp...

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Problems solved by technology

[0004] 1) Because cell lines are easy to culture, economical and efficient, tumor models established with cell lines such as U251 and 9L are relatively common. For example, Chinese Patent No. CN200610000881.2 discloses a 9L that stably expresses firefly luciferase LUC The construction and application of cell lines, but because the cell lines are continuously passaged in vitro, the continuously passaged cell lines adapt to the environment of the external culture dish, gradually lose the heterogeneity of the tumor and the related signaling pathways also change, so it cannot accurately represent Physiological changes of cells in the body

[0005] 2) The tumor model based on inbred immunodeficient mice that can stably express green fluorescent protein (GFP) can intuitively distinguish the complex relationship between the tumor and the host, as disclosed in Chinese Patent No. CN201710452157.1. The method and model application of green fluorescent BALB / c nude mouse model, but GFP has strong background interference, its sensitivity and accuracy are greatly reduced, and the cycle of cultivating suitable mice is too long, costly and complicated to operate

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