The development of animal model for chromosome trisomy-syndrome by microcell-mediated chromosome transfer

微细胞介导染色体转移建立染色体三体综合征动物模型

• 批准号: 09480247
• 负责人: FUNAKI Kenji
• 金额: $ 2.82万
• 依托单位: Shimane University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09480247/
• 关键词: microcell-mediated chromosome transfer; human chromosome 21; mouse embryonic stem cell; chimeric mouse; キメラマウス; ヒト染色体; 胚幹細胞; ES細胞; マウス; キメラ

In present study, hChr.21 was introduced into mouse embryonic stem (ES) cells via microcell-mediated chromosome transfer (MMCT), and viable chimeric mice were produced from them. We analyzed theses mice as followings. Metaphase spreads of various tissues from four chimeric mice were examined by Fluorescence in situ hybridization (FISH) to evaluate the stability of hChr.21 in somatic cells. The percentage of the spreads containing the hChr.21 showed 50-100%. RT-PCR analysis of one of these mice showed that several genes on hChr.21 were expressed in a proper tissue-specific manner. Finally, we have determined that chimeric mice could transmit hChr.21 fragment (hChr.21f) to their offspring through germline. FISH analysis of various tissues in these germline mice showed that the retention of hChr.21f of them were lower than that of chimeric mice.In anatomical and histological observation of 11 chimeric mice dead in perinatal period, it was found that aplasia of the sternum and remarkable decrease of germ cell occurred in all of them, and various histological abnormality occurred sporadically in heart, liver and kidney. As a control, four chimeric mice induced hChr.2f (containing immunoglobulin genes) were anatomically and histologically studied. In almost of them delayed ossification of the cervical vertebra centrum, aplasia of the sternum and polydactyly of the forelimb were observed, and hypertrophy and histologically various abnormality in hert, liver and kidney occurred sporadically. It is highly possible that remarkable decrease of germ cell in the hChr.21-chimeric mice and polydactyly in the hChr.2f-chimeric mice are related to the gene(s) on the human chromosome introduced.

本研究通过微细胞介导的染色体转移（MMCT）将hChr.21导入小鼠胚胎干细胞（ES细胞），并从它们产生活的嵌合小鼠。我们对这些小鼠进行了如下分析。通过荧光原位杂交（FISH）检查来自四只嵌合小鼠的各种组织的中期扩散，以评估hChr.21在体细胞中的稳定性。含有hChr.21的涂抹片的百分比显示为50- 100%。对其中一只小鼠的RT-PCR分析表明，hChr.21上的几个基因以适当的组织特异性方式表达。最后，我们确定嵌合小鼠可以通过生殖细胞将hChr.21片段（hChr.21f）传递给后代。对11只围产期死亡的嵌合体小鼠进行解剖学和组织学观察，发现其胸骨均发育不全，生殖细胞明显减少，心、肝、肾等组织学均有不同程度的异常。作为对照，4只嵌合小鼠诱导hChr.2f（含有免疫球蛋白基因）的解剖学和组织学研究。大多数病例颈椎椎体骨化延迟，胸骨发育不全，前肢多指畸形，偶有肥大及肝、肾等组织学异常。hChr. 21-嵌合小鼠中生殖细胞的显著减少和hChr. 2f-嵌合小鼠中的多指畸形很可能与引入的人染色体上的基因有关。

项目成果

K. Tomizuka: "Double trans-chromosomic mice : Maintenance two individual human chromosome fragments containing Ig heavy and κ loci"Proc. Natl. Acad. Sci. USA. 97. 722-727 (2000)

K. Tomizuka：“双转染色体小鼠：含有 Ig 重链和 κ 基因座的两个个体人类染色体片段”Proc. Natl. 97. 722-727 (2000)。

M. Oshimura: "Mice with introduction of human chromosome"Experimental Medicine. 16. 511-514 (1998)

M. Oshimura：“引入人类染色体的小鼠”实验医学。

K. Tomizuka: "Functional expression and germline transmission of a human chromosome fragment in chimeric mice"Nature Genet.. 16. 133-143 (1997)

K. Tomizuka：“嵌合小鼠中人类染色体片段的功能表达和种系传递”Nature Genet.. 16. 133-143 (1997)

K.Tomizuka: "Double trans-chromosomic mice : Maintenance of two individual human chromosome fragments containing Ig heavyand κ lici."Proc.Natl.Acad.Sci.USA. 97. 722-727 (2000)

K.Tomizuka：“双转染色体小鼠：含有重链 Ig 和 κ lici 的两个个体人类染色体片段的维持。”Proc.Natl.Acad.Sci.USA 97. 722-727 (2000)。