摘要
目的:建立一个能够稳定传代且生物学特性稳定的兔舌癌细胞系并对其生物学特性进行检测。方法:取对数生长期的第55代RSCC-1,注射于新西兰白兔舌缘,2周后成瘤,取瘤体组织Transwell Permeable Supports(TPS)法原代培养、纯化,获得新的兔舌癌细胞系RSCC-1a。连续传代培养,并对RSCC-1a进行生物学特性检测:1.形态学检测,包括HE染色、免疫组化染色、倒置相差显微镜、透射电镜观察兔舌癌细胞形态;2.细胞生长动力学检测,包括生长曲线绘制、细胞生长周期分析、计算细胞倍增时间;3.成瘤性检测,包括同种异体(兔)成瘤率、异种(裸鼠)成瘤率;4.核型分析。结果:RSCC-1接种后2周,形成兔舌癌模型,病理检查证实为鳞状细胞癌,瘤体组织进行原代培养,获得RSCC-1a细胞系,已稳定传至73代,形态学观察见细胞呈多边形,免疫组化角蛋白阳性。生长曲线呈停滞期、对数生长期及平台期3个典型时相;细胞周期测G1期为71.55%,G2期为6.59%,S期为20.84%;染色体为亚四倍体核型,染色体数为93~212条,众数为139;裸鼠成瘤率100%,同种异体成瘤率100%,部分兔还发生颈部淋巴结转移。结论:通过RSCC-1成瘤后的瘤体组织进行原代培养,获得兔舌癌细胞系RSCC-1a,对其生物学特性检测,证明其为上皮来源的恶性肿瘤细胞系。
Objective:To establish a stable passage and stable biological characteristics of rabbit tongue cancer cell line and to detect its biological characteristics.Method:RSCC-1 of the 55 th generation in the logarithmic growth stage was injected into the lingual margin of New Zealand white rabbits,and after 2 weeks,the tumor was formed.The primary culture of the tumor tissues with Transwell Permeable Supports(TPS)was performed to obtain a new rabbit tongue cell line RSCC-1 a after purification.Continuously subculture and detect the biological characteristics of the new cell lines:1.detection of morphological,including HE staining,Immunohistochemistry staining and using inverted phase contrast microscope,to observe cell morphology.2.detection of cell growth kinetics,including growth curve drawing and cell cycle analysis and cell doubling time calculation.3.detection of tumor,including allogeneic tumor formation rate(rabbit)and xenograft tumor formation rate(mouth).4.Karyotype analysis.Results:Two weeks after the inoculation of RSCC-1,a rabbit tongue cancer model was formed,which was confirmed to be squamous cell carcinoma by pathological examination.The RSCC-1 a cell line was obtained by primary culture of the tumor tissue,which has been transferred to 73 generations steadily.Growth curve shows three typical phases,respectively,stagnation,logarithmic phase and plateau.Measure the cell in cell cycle can find that G1 phase is71.55%,G2 is 6.59%and S phase is 20.84%.Chromosome is the karyotype of tetraploid.The number of chromosomes is 93 to 212 and the plural is 139.Nude mice tumor rate is 100%,allogeneic tumor formation rate is 100%.Cervical lymphatic metastasis also occurred in some rabbits.Conclusion:Get new cell lines by primary culturing tumor tissue that forming of RSCC-1.Detecting its biological characteristics proves that it is a squamous epithelioma from epithelium.
作者
于化蛟
王姝
金武龙
YU Hua-jiao;WANG Shu;JIN Wu-long(Affiliated Hospital of Inner Mongolia Medical University,Inner Mongolia 010050,China)
出处
《中华老年口腔医学杂志》
2020年第3期146-150,155,共6页
Chinese Journal of Geriatric Dentistry
基金
内蒙古自治区自然科学基金(项目编号:2019MS08141)
关键词
兔舌癌
细胞培养
鳞状细胞癌
细胞系
rabbit tongue cancer
cell culture
cell line
biological characteristics
