| Co-Investigator(Kenkyū-buntansha) |
YOSHIDA Mitsuaki A. Res. Assoc., Med. Res. Inst., Tokyo Med. Dent. Univ., 難治疾患研究所, 助手 (60182789)
YAMAMOTO Kohtaro Assoc. Prof., Med. Res. Inst., Tokyo Med. Dent. Univ., 難治疾患研究所, 助教授 (40000971)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Research Abstract |
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
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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
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