人肝癌细胞系HC-108的建立及其生物学特征

HC108细胞系为贴壁生长具有上皮细胞形态特征的分化程度较高的人肝癌细胞。AFP分泌阴性,细胞平均倍增时间为32.21小时,细胞分裂指数为27.75‰,软琼脂克隆形成率为5.0％,能形成裸鼠移植瘤,支原体为阴性。染色体和流式细胞术分析为超二倍体核型,17号染色体改变最为明显。     

家蝇胚胎细胞系的建立及其生物学特性

昆虫细胞系在许多领域得到广泛的应用。以家蝇Musca domestica L.卵(胚胎)为材料,用含20%胎牛血清的M3培养基培养,原代培养35d左右形成单层,每7d传代1次,连续传代60代次。细胞群体倍增时间为44h,染色体2n=12,可在液氮及4℃冰箱中保存复苏,命名为MDEC-07114。     

人成纤维细胞系HF—91的建立及生物学特性

人胚胎胰腺结缔组织经分离培养成人成纤维细胞系ＨＦ－９１,传４０代。该细胞贴壁生长,具有密度依赖性抑制性质。它的生长依赖于成纤维细胞生长因子的存在,在１０ｎｇ／１００ｎｇ／ｍｌ浓度范围内,细胞生长与ｂＦＧＦ的浓度成正相关。细胞群体倍增时间２３．８±５．３ｈ。有丝分裂指数４９．３％±４．１％染色体众数２ｎ＝４６,占８７．５％±４．１％。染色体众数２ｎ＝４６,占８７．５％－９１．０％,乳酸脱氢酮同工酶Ｌ     

云南宣威肺腺癌细胞系SLC—89的建立及其生物学特性

本文报告1例来源于云南宣威县患者的肺癌标本,经体外培养建系成功,命名为SLC-89。该细胞系细胞经HE,瑞氏染色形态符合癌细胞特征。在体外培养已两年多,传代196代,细胞冻存后复苏生长良好。第86代细胞倍增时间为26.4小时,染色体数为非整倍体,众数为超二倍体,长期培养后染色体数明显增加。细胞接种裸鼠有移植瘤生长,组织象与原发肺癌组织象相似。电镜观察细胞表面有微绒毛,浆中可见分泌颗粒,有较多板层小体,表明来源于肺泡上皮。     

锦鲤鳍条组织细胞系的建立及其生物学特性

采用组织块移植培养技术,对来源于锦鲤（Cryprinus carpiod）鳍条组织的细胞进行原代培养,建立了锦鲤鳍条组织细胞系,已稳定传代60多次,命名为Koi-Fin。锦鲤鳍条组织细胞为成纤维样细胞,最佳培养基为MEME,最适血清体积分数为10%,最适培养温度为25 oC,群体倍增时间为43.5 h。该细胞经液氮冷冻保藏12个月后采用台盼蓝染色,约（80.21±5.84）%的细胞具有细胞活性,复苏细胞生长旺盛。细胞染色体分析显示,第16代锦鲤鳍条组织细胞的染色体数目为正常二倍体2n=100,第40代细胞的染色体众数为52。病毒敏感性试验结果表明,Koi-Fin细胞系对锦鲤疱疹病毒（Koi Herpesvirus,KHV）敏感,可产生典型细胞病变效应,病毒滴度为107.86±0.51TCID50/mL。针对锦鲤疱疹病毒胸苷激酶（thymidine kinase,TK）基因设计特异性引物进行PCR检测,可扩增出病毒靶基因片段。     

人黑色素瘤细胞系（HMM—239）的建立及其生物学特性

人黑色素瘤细胞系在国外虽已建立近200个[1],但在国内尚未见报道。我们于1987年首次将一活检病理诊断为左足趾黑色瘤的患     

中国鄂温克、鄂伦春、达斡尔族永生细胞系的建立与保存

本文采用EBV（Epstein?Barr Virus）上清液转化B淋巴细胞,并加入环胞霉素A（Cyclosporine A）抑制T淋巴细胞,成功地对中国东北地区鄂温克族、鄂伦春族及达斡尔族的部分个体建立了永生细胞系,其中鄂温克族49株,鄂伦春族40株,达斡尔族51株,总计140株。永久保存我国特有民族的基因组,为分析其遗传学差异奠定了基础。 Abstract：The immortal lymphoblastoid cell lines were established by EBV transformation of B cells and addition of cyclosporin A to inhabit the activity of T cells.In the present study ,140 immortal cell lines of the Ewenki,the Oroqen and the Daur ethnic groups in the Northeast China were established .This is an important part of the research of human genome diversity for the exploration of the origin and progression of different ethnic groups ,and also provide enough research materials for further studies.     

中华按蚊（Anopheles sinensis）细胞系的建立和特征

从中华按蚊初孵幼虫中,建立一株细胞系,命名 AS-684。建系使用我们改良的199培养基及含有10%F 12的 M-M 培养基。原代培养20天形成单层细胞。传代培养,以1:2分种率,相隔4—5天传代1次,连传70代次。细胞系由4种细胞类型组成,以多边形和纺锤形上皮细胞为优势,掺杂少数双核细胞及长梭形细胞。细胞生长,从24小时开始倍增分裂达高峰,对数期二倍增长时间约为16小时。自45代起,培养基中血清含量从20%降至10—15%。4次电镜观察,没有发现支原体和病毒质点。细胞系以二倍体细胞为主,染色体6条(2n=6)。细胞酯酶同工酶分析,酶谱为两条酶带,PI 值为4.75,4.85。各种代次细胞保存于-196℃的液态氮中及4℃普通冰箱中,经多次试验能成功复苏。     

云南矮马耳缘组织成纤维细胞系的建立及其生物学特性

随着全球生物多样性及其遗传资源丧失程度的加剧 ,使得各国纷纷投入大量人力和财力 ,并以多种方式收集、保存和整理大自然赋予人类的宝贵财富 ,如美国典型培养物保藏中心 (AmericanTypeCultureCollection ,ATCC) ,欧洲动物细胞培养物保藏中心 (EuropeanCollectionofAnimalCellC     

高转移潜力食管癌细胞株的建立及其生物学特征

目的建立具有高转移潜力食管癌细胞株并研究其生物学特征。方法将食管癌细胞系EC109细胞悬液异位移植到SCID小鼠胃壁,约3个月后或动物濒临死亡时处死,行病理学解剖,将肉眼可见的纵隔淋巴结转移瘤块接种于SCID鼠皮下扩增,然后取小鼠皮下瘤组织块进行细胞培养,得到性状稳定的细胞株NMC109后,用MTT法分析细胞生长曲线,Western bloting法检测与细胞分裂增殖能力密切相关的TopoⅡα表达,酶谱法检测MMP-2和MMP-9的活性,划痕实验和Transwell体外移动实验检测细胞的移动能力。结果与母本细胞EC109相比,所获得的细胞株NMC109其增殖能力和TopoⅡα表达明显增强,MMP-9的活性明显升高,移动能力明显增强。结论获得了具有高转移潜力的食管癌细胞株。     

无线粒体DNA的人肺腺癌A549细胞系的构建

目的:建立无线粒体DNA(mtDNA)的人肺腺癌ρ~0A549细胞系。方法:在含50 ng/mL溴化乙锭(EB)、100μg/mL丙酮酸钠和50μg/mL尿嘧啶核苷的RPMI1640细胞培养基中传代培养A549细胞;用低剂量EB连续诱导培养35 d后,采用光镜观察、TaqMan探针法实时荧光定量PCR(qPCR)和Western印迹鉴定无mtDNA的ρ~0A549细胞系;采用MTT法测定ρ~0A549细胞增殖曲线。结果:倒置显微镜下野生型ρ^+A549细胞为多角形,ρ~0A549细胞形态呈拉长枝状;qPCR结果显示,低剂量EB诱导35 d的ρ~0A549细胞中无mtDNA的存在。Western印迹结果显示,ρ^+A549细胞中能表达核基因编码的线粒体蛋白SDHA和ATP5A,也能表达线粒体基因组编码的蛋白MT-COXI和MT-ATP6;ρ~0A549细胞中无MT-COXI和MT-ATP6蛋白表达,但核基因编码的SDHA和ATP5A蛋白能够正常表达。MTT结果显示,与ρ^+A549细胞相比,ρ~0A549细胞生长速度明显减慢,差异有统计学意义(P<0.05)。结论:构建和鉴定了无mtDNA的人肺腺癌ρ~0A549细胞系,为后续探讨mtDNA缺失或突变与人肺腺癌发生之间的关系奠定了实验基础。     

A549人肺癌细胞系/615-SCID小鼠转移瘤的生物学特征

目的通过建立A549人肺腺癌细胞/615-SCID小鼠模型,评价重度联合免疫缺陷615-SCID小鼠在建立人类肺癌转移模型方面的应用价值.方法将1×107A549细胞接种到615-SCID及SCID小鼠右上肢背部皮下,观察成瘤时间、成瘤率、肿瘤生长速度及转移发生.结果两品系小鼠接种后的成瘤率均为100%,615-SCID小鼠移植瘤潜伏期较长、生长较缓慢,更容易发生转移.结论 615-SCID小鼠比SCID小鼠更易于构建人类肺腺癌转移模型,对于肺癌转移特性研究具有较大的意义.     

鼻咽癌CD44抑制表达细胞株的建立

目的:构建CD44新剪接变异体siRNA质粒表达载体,建立CD44新变异体抑制表达的鼻咽癌细胞株.方法:合成CD44新变异体特异性干扰DNA片段,干扰DNA片段亚克隆于带绿色荧光蛋白的pGenesil -1.3质粒表达载体中,双酶切和测序鉴定重组表达质粒载体;采用脂质体将重组表达质粒载体转染入鼻咽癌5 -8F细胞系,进行G418筛选;Western blot分析CD44表达.结果:重组CD44干扰DNA片段质粒表达载体的碱基序列和插入方向正确;细胞转染效率达70％;G418筛选获得GFP表达的单克隆鼻咽癌5 -8F细胞株;Western blot分析表明CD44表达受抑制.结论:建立了CD44新变异体抑制表达的鼻咽癌细胞株,为CD44新变异体在鼻咽癌中的生物学功能提供了基础.     

Establishment and Partial Characterization of a Continuous Human Malignant Glioma Cell Line: NG97

1. A human glioma cell line, NG97, was established from tissue obtained from a patient diagnosed with a grade III astrocytoma.2. The NG97 cell line has been subcultured for more than 100 passages in standard culture media without feeder layer or collagen coatings.3. NG97 cells grow in vitro as two subpopulations with distinct morphological appearance: stellate cells with pleomorphic nuclei, and small round cells with few processes. The cells have a doubling time of about 72 h and a plating efficiency of 1%. The injection of NG97 cells into congenitally athymic mice induced the formation of solid tumor masses that could be retransplanted every 4 weeks. The cells obtained from tumor mass when cultivated in vitro had a morphology comparable to those of the initial culture.4. This cell line may prove useful for cellular and molecular studies as well as in studies of gliomas treatment.     

乙型肝炎病毒和黄曲霉素协同作用诱发人肝细胞癌细胞株的建立

为了研究人乙型肝炎病毒（HBV）和黄曲霉素（AFB1）在肝癌发生过程中的作用,我们用HBV感染的人胚胎肝细胞移植至裸鼠背部皮下,以后每周皮下注射AFB1,在裸鼠体内成功地诱发了人肝细胞癌。选3只裸鼠所形成的肿瘤组织,分别再接种裸鼠传代。在裸鼠体内传至5代后,将瘤组织在体外培养、传代,建立了3个肝癌细胞株,分别为CBH－1a、CBH－1b和CBH－2。对3个细胞株进行生物学特性分析发现,细胞生长迅速,接触抑制消失,细胞增殖核主数Ki67阳性细胞占38．2％,用EMA单抗检测证实为人来源细胞,核酸原位杂交显示,细胞中HBV－X和HBV－S基因阳性,PCR可扩增出X基因,证明HBV基因已到瘤细胞中,3个细胞株细胞再接种裸鼠皮下,可再生成肿瘤,此实验证明了HBV协同AFB1在个肝细胞癌发生过程中的病因作用。同时,为进一步研究HBV和AFB1在肝癌发生过程中的分子机制提供了细胞水平的模型。     

Isolation and Characterization of a Near-Haploid Human Cell Line

Mammalian somatic cells are usually diploid. Occasional rare human tumors have been shown to have a hypodiploid karyotype. We have isolated a near-haploid subclone (P1-55) from a heterogeneous human leukemia cell line, KBM-7. These near-haploid cells have approximately half the human diploid DNA content and have a haploid karyotype except for a disomy of chromosome 8 (25, XY, +8, Ph(+)). This cell line maintains a majority of cells with a near-haploid karyotype for at least 12 weeks in culture. By serial subcloning, we have isolated near-haploid subclones that maintain ploidy for at least 8 months in culture. Near-haploid cells can also be efficiently isolated from mixed ploidy cultures by size selection. The availability of this human near-haploid cell line should facilitate the genetic analysis of cultured human cells.     

人肺腺癌细胞A—549和正常细胞HBE的蛋白质组差异分析

为了研究人肺腺癌细胞A 5 49和正常细胞HBE的蛋白质组差异 ,用固相pH梯度双向凝胶电泳分离人肺腺癌细胞系A 5 49和正常细胞HBE的总蛋白质 ,银染显色 ,PDQuest 2 DE软件分析 ,对部分差异蛋白质点进行基质辅助激光解吸电离飞行时间质谱 (MALDI TOF MS)测定其胶内酶解后的肽质指纹图谱 ,用PeptIdent软件查询SWISS PROT数据库。结果获得了分辨率和重复性均较好的双向电泳银染图谱 ,图象分析探测到A 5 492 DE图谱的平均蛋白质点数为 (890± 38)个 ,HBE的平均蛋白质点数为 (75 7± 2 7)个 ,不同胶间蛋白质点的位置偏差在IEF方向为 (2 .85± 0 .48)mm ,在SDS PAGE方向为 (2 .6 9± 0 .37)mm。差异表达分析发现A 5 49和HBE图谱有5 35个蛋白质点相互匹配 ,其中A 5 49有 35 5个未被匹配 ,HBE中有 2 2 2个未被匹配 ;对A 5 49和HBE中的 18个差异蛋白质点分别进行肽质指纹分析 ,经数据库查询 ,初步鉴定为一些与物质代谢、细胞因子、信号转导有关的蛋白质。提示人肺腺癌细胞A 5 49和正常细胞HBE的蛋白质组具有差异 ,这种蛋白质组的差异分析有助于进一步研究肺腺癌的相关蛋白质及分子标记物     

Establishment of a New Human Glioblastoma Multiforme Cell Line (WJ1) and Its Partial Characterization

(1) A new human glioblastoma multiforme (GBM) cell line, WJ1, was established from the tissue derived from a 29-year-old patient diagnosed with a grade IV GBM. (2) The WJ1 cell line has been subcultured for more than 80 passages in standard culture media without feeder layer or collagen coatings. (3) GBM cells grow in vitro with distinct morphological appearance. Ultrastructural examination revealed large irregular nuclei and pseudo-inclusion bodies in nuclei. The cytoplasm contained numerous immature organelles and a few glia filaments. Growth kinetic studies demonstrated an approximate population doubling time of 60 h and a colony forming efficiency of 4.04%. The karyotype of the cells was hyperdiploid, with a large subpopulation of polyploid cells. Drug sensitivities of DDP, VP-16, tanshinone IIA of this cell line were assayed. They showed a dose- and time-dependent growth inhibition effect on the cells. (4) Orthotopic transplantation of GBM cells into athymic nude mice induced the formation of solid tumor masses about 6 weeks. The cells obtained from mouse tumor masses when cultivated in vitro had the same morphology and ultrastructure as those of the initial cultures. (5) This cell line may provide a useful model in vitro and in vivo in the cellular and molecular studies as well as in testing novel therapies for human glioblastoma multiforme.     

Establishment and Characterization of a Nontumorigenic Cell Line Derived from a Human Hepatocellular Adenoma Expressing Hepatocyte-Specific Markers

In the present study the establishment and characterization of a nontumorigenic liver epithelial cell line (HACL-1) derived from a human hepatocellular adenoma is described. The HACL-1 cells have a finite life span (i.e., they proliferate for a period of 2 months and then senesce), show cell–cell contact inhibition, do not grow in soft agar, are not tumorigenic when injected in nude mice, and possess a normal diploid karyotype. The cultured cells resemble hepatocytes, but exhibit some features of dedifferentiation. At the ultrastructural level the cells are endowed with round or oval nuclei, abundant cytoplasmic organelles, and varying amounts of glycogen. The rough endoplasmic reticulum is disorganized, while peroxisomes and matrix granules within mitochondria are lacking. HACL-1 cells are cytokeratin 18-positive as well as (transiently) albumin- and α-fetoprotein-positive, but do not express cytokeratin 19. Furthermore, no mutations were observed in exons 5–8 of the tumor suppressor genep53.Taken together these results show that HACL-1 cells are nontumorigenic proliferating liver epithelial cells, which might prove to be of great value in future studies on diverse aspects of human liver cell biology and carcinogenesis.