摘要
目的探讨细胞同步化染色体高分辨技术检测杂交瘤细胞染色体的可行性。方法选择小鼠骨髓瘤SP-2/O细胞株、人淋巴母细胞CKM-8细胞株、人骨髓瘤KM细胞株、抗小鼠胰腺癌鼠-鼠杂交瘤细胞株,体外培养24～48h。对4种细胞株分别进行同步化处理,加入5-氟脱氧尿嘧啶核苷和尿嘧啶培养16～18h后,加入5-溴脱氧尿嘧啶核苷,并在收集细胞前加入秋水酰胺;收集细胞进行离心、低渗处理、固定、再固定,制备染色体滴片,应用胰蛋白酶-Giemsa染液对染色体进行G显带处理。每例标本以30个以上分裂象做众数分析,按照《人类细胞遗传学国际命名体制》(ISCN 1978)对细胞株染色体进行G显带分析。结果 4种细胞株的染色体均较长、分散良好、形态较好。SP-2/O细胞株、CKM-8细胞株、KM细胞株、抗小鼠胰腺癌鼠-鼠杂交瘤细胞株的染色体数目分别为63、46、42、105,其中抗小鼠胰腺癌鼠-鼠杂交瘤细胞染色体数目接近小鼠淋巴细胞和小鼠骨髓瘤细胞染色体数的总和,且多数为端着丝点染色体。结论细胞同步化染色体高分辨技术检测杂交瘤细胞染色体具有良好的效果。
Objective To investigate the feasibility of detecting hybridoma chromosome by using synchronized chromosome separation technique.Methods Mouse myeloma cell line SP-2/O cells, human lymphoblastoid cell line CKM-8 cells, human myeloma cell line KM ceils, and mouse-mouse hybridoma （anti-mouse pancreatic cancer） cells were cultured for 24-48 hours in vitro. Chromosomal synchronization was carried out in four cell lines above by treating cells with 5-fluoro-deoxy uridine and uracil for 16-18 hours, then adding 5-BrdU into culture. Cells were treated by Colcemid CCM before harvesting. After centrifugation, cells were treated by hypotonic buffer and followed by double-fix procedures. The liquid form of chromosome pieces were dropped onto glass slides and G-banding was carried out by staining slides with trypsin-Giemsa solution. Mode analysis was performed by using more than 30 separation phase of chromosomes in each case. Chromosomal G-banding analysis was carried out according to ＂Human Cytogenetics International Naming System＂ （ISCN 1978）.Results The chromosomes obtained from four cell lines had ideal length with good separation and clear shape. The chromosomal numbers of SP-2/O cells, CKM-8 cells, KM cells and mouse-mouse hybridoma （anti-mouse pancreatic cancer） cells were 63, 46, 42 and 105 respectively. The numbers of chromosomes of the mouse-mouse hybridoma （anti-mouse pancreatic cancer） cells was close to sum of the numbers of chromosomes of the mouse spleen cells and mouse myeloma cells, and most of the chromosomes of mouse-mouse hybridoma （anti-mouse pancreatic cancer） cells were telocentric chromosome.Conclusion Synchronized chromosome separation technique was effective in detecting hybridoma cells chromosome.
出处
《中国医药生物技术》
CSCD
2010年第5期367-370,共4页
Chinese Medicinal Biotechnology
关键词
杂交瘤
细胞遗传学分析
染色体
动物实验
Hybridomas
Cytogenetic analysis
Chromosomes
Animal experimentation
