Chromosomal Instability in Lymphoblastoid Cell Lines Derived from Patients with Different Inherited disorders

不同遗传性疾病患者来源的淋巴母细胞系的染色体不稳定性

基本信息

批准号：
61571089
负责人：
IKEUCHI Tatsuro
金额：
$ 1.41万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

项目摘要

Permanent growing lymphoblastoid cell lines (LCL) established by EB virus-mediated transformation of lymphocytes are now of great practial value in both clinical and experimental human genetics. We have established a series of LCL from patitents or families with various hereditary diseases. In order to confirm their availability and to obtain the basic guidelines for the maintenance of LCL, we have examined cytogenetic characters of these LCL in the course of prolonged culture condition. The results are as follows:1) The LCL established for the last 2 years include: 87 lines from 36 families with familial polyposis coli (FPC), 24 lines from 6 families with multiple endocrine neoplasia type 2 (MEN2), 19 lines from heritable fragile site (FS) carriers, and 14 lines from patients with various chromosomal abnormalities.2) Karyotype analyses were made on prolonged cultures (2 months to 1.5 year) of LCL from MEN2 families and from FS carriers. In general,acquired chromosome abnormalities app … More eared four months after the establishment, regardless of their genetic sources (age, ses, carriers or noncarriers). In some of the LCL maintained in vitro for more than one year, almost all of the cells showed abnormal karyotypes with clonal chromosome changes. The abnormalities were mostly of numerical type with a predominance of trisomies of Nos. 5, 8, 12 and 15 chromosomes.3) Expression rates of FS in LCL derived from heritable FS carriers were in general very low (<3%), but a cell line "B-3",derived froma RdU-required fra(10)(q25) carrier, expressed the FS with high frequencies (40-60%) after exposure to 5-bromodeoxyuridine (7<micrn>g/ml) for 24 hours. 4) In LCL from patients with a ring chromosome, r(18) and r(21), respectively, the rings were retained even after prolonged culture for 4 months. This indicates the morphological stability of the rings in which the small-sized chromosomes were involved.5) Some of the LCL here establsihed were efficiently employed for regional mapping of cloned DNA segments (D13S21), D13S22 and D18S5). The LCL from MEN2 and FPC families could be applied to the linkage analysis and to the esarch inn tumors for somatic loss of constitutinal heterozygosity.6) In experiments using the cell line "B-3", it was found that the fra(10)(p25) expression distribution of thymine between the two strands of DNA duplex in the fra(10) site. Less

EB病毒介导的淋巴细胞转化建立的永久性生长淋巴母细胞系（LCL）现在在临床和实验性人遗传学中都具有巨大的实践价值。我们已经从患有各种遗传性疾病的患者或家庭建立了一系列LCL。为了确认其可用性并获得维持LCL的基本准则，我们在延长培养条件的过程中检查了这些LCL的细胞遗传学特征。结果如下：1）过去2年建立的LCL包括：来自36个家庭息肉病大肠杆菌（FPC）的87条线，来自6个患有多个内分泌肿瘤2（MEN2）的6个家庭的24行，来自可遗传的脆弱部位（FS）载体的19条线，以及来自各种染色体abnormal abnormal and the watermality的患者。 Men2家族和FS运营商的LCL年。通常，收购的染色体异常应用程序……在建立四个月后，更担心，无论其遗传来源如何（年龄，SES，载体或非载体）。在某些LCL在体外维持一年以上，几乎所有细胞均显示出异常的核型，具有克隆染色体的变化。异常性主要是数值类型，trisomies占5、8、8、12和15号染色体的占主导地位。3）fs的LCl表达速率。源自赫洛可携带的FS载体的表达速率通常非常低（<3％），但源自RDU-3频率（<3％），是rdu-requient fra fra fra（q25）（Q25）（Q25）（Q25）（Q25）（Q25）（Q25）（40-60％）暴露于5-溴脱氧尿苷（7 <mincn> g/ml）24小时后。 4）在有环染色体的患者的LCL中，R（18）和R（21）分别在长时间培养后仍保留环。这表明涉及小型染色体的环的形态稳定性。5）5）这里建立的一些LCL有效地用于克隆DNA片段的区域映射（D13S21），D13S22），D13S22和D18S5）。 MEN2和FPC家族的LCL可以应用于链接分析和Esarch Inn肿瘤，以造成构造杂合性的体细胞丧失。6）在实验中使用细胞系“ B-3”，发现FRA（10）（P25）在fra（10）dna Duplex in fra（10）中的胸腺表达分布（P25）。较少的