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胚胎干细胞(embryonic stem cells,ESCs)是从囊胚的内细胞团分离出来的多潜能干细胞,具有多向分化的能力。将外源基因导入ES细胞建立转基因动物,对于研究外源基因的功能和调控具有一定的价值。载有外源性基因的病毒在感染ES细胞后,可通过囊胚注射获得具有胚系遗传的该转基因动物,并且这一外源基因可以稳定遗传和表达。该研究主要是利用携带hPML-RARα基因的慢病毒感染小鼠ES细胞系(R1),获得携带该基因的ES细胞,感染后的ES细胞核型正常。在此基础上,将感染后的ES细胞经囊胚注射,获得了携带有hPML-RARα基因的3只嵌合小鼠,其中,有1只具有遗传特性。对嵌合体小鼠与C57杂交的后代给予强力霉素(doxycycline)处理,3天以后骨髓细胞hPML-RARα基因开始表达,这证明了在小鼠体内该外源基因表达的可诱导性。以上证实,已经成功利用ES细胞建立了可诱导的白血病转基因小鼠模型。
Embryonic stem cells (ESCs) are pluripotent stem cells isolated from the inner cell mass of the blastocyst and have the ability to differentiate into multiple directions. The introduction of exogenous genes into ES cells to establish transgenic animals is of value in studying the function and regulation of exogenous genes. The virus carrying the exogenous gene can infect ES cells, and the transgenic animal with germline inheritance can be obtained by blastocyst injection, and the foreign gene can be stably inherited and expressed. In this study, ES cells carrying this gene were obtained by inoculating the mouse ES cell line (R1) with a lentivirus carrying the hPML-RARα gene. The infected ES cells were normal in karyotype. Based on this, 3 infected chimeric mice carrying hPML-RARα gene were obtained by blastocyst injection of infected ES cells, of which 1 had genetic characteristics. Administration of doxycycline to progeny of chimeric mice that hybridized with C57 began 3 days after the expression of the hPML-RARα gene began to be expressed in the bone marrow cells, demonstrating the inducibility of the exogenous gene expression in mice. The above confirmed that ES cells have been successfully used to establish an inducible leukemia transgenic mouse model.