Establishment of an animal model for human cellular againg

人类细胞再次动物模型的建立

• 批准号: 11554038
• 负责人: ISHIDA Takafumi
• 金额: $ 5.12万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11554038/
• 关键词: macaque; cellular aging; in vitro; human; model; 細胞培養; 老化; p53; ヘリケース; エイジング; 細胞老化; 細胞株化; テロメア

To clarify the pattern of macaque cellular aging, a total of 40 cultures of Japanese macaque (Macaca fuscata), long-tailed macaque (Macaca fascicularis), bonnet monkey (Macaca radiata) and rhesus macaque (Macacamulatta) were carried out. Adherent cells were isolated from skin, kidney and lung, cultured in RPMI 1640 medium supplemented with 10% FBS, and subcultured until they terminated cell division. In 36 cultures, cells underwent senescence showing lager cell size and decreased saturation density, and then terminated cell division (Mortality stage 1 (M1)) at 10-20 Population Doubling Levels (PDLs). Whereas cells in four cultures first exhibited senescent morphology at 15-25 PDLs, but continued cell division through M1 up to 40-100 PDLs and then came into crisis (Mortality stage 2 (M2)). Cells in one culture of Japanese macaque do not terminate cell division (>150 PDLs) and showed several characteristics of transformed cells, such as the anchorage independent growth and loss of contact inhibition. Shortening of telomeres by PDLs and the absence of telomerase activity were identified in all of tested including the transformed one. Macaque cells showed an intermediate pattern of in vitro aging between human and rodent cells, whereas, in terms of telomerase activity, they are relatively close to human cells. It is thus suggested that the macaques serve as animal models for human cellular aging and that they provide us with a key to study possible mechanisms which play roles in the critical stages (M1 and M2) in cellular aging and transformation.

为了阐明猕猴细胞衰老的模式，对日本猕猴（Macaca fuscata）、长尾猕猴（Macaca fascicularis）、帽猴（Macaca radiata）和恒河猴（Macacamulatta）共40只进行了培养。从皮肤、肾和肺分离贴壁细胞，在补充有10%FBS的RPMI 1640培养基中培养，并传代培养直至它们终止细胞分裂。在36个培养物中，细胞经历衰老，显示较大的细胞尺寸和降低的饱和密度，然后在10-20个群体倍增水平（PDL）时终止细胞分裂（死亡阶段1（M1））。而四种培养物中的细胞在15-25个PDL时首先表现出衰老形态，但通过M1继续细胞分裂直至40-100个PDL，然后进入危象（死亡阶段2（M2））。在日本猕猴的一种培养物中的细胞不终止细胞分裂（>150个PDL），并且显示转化细胞的几个特征，例如不依赖于锚定生长和失去接触抑制。在所有的测试中，包括转化的一个，PDLs的端粒缩短和端粒酶活性的情况下进行了鉴定。猕猴细胞在体外老化过程中表现出介于人类和啮齿动物细胞之间的中间模式，而在端粒酶活性方面，它们相对接近人类细胞。提示猕猴可作为人类细胞衰老的动物模型，为研究细胞衰老和转化的关键阶段（M1和M2）可能的机制提供了一把钥匙。

项目成果

Suzuki, J., Ishida, T., Ohkura, S., Udono, T., Hayasaka, I.and Araithamkul, V.: "Plasma IGF-1 levels in the great and lesser apes."Proc.XVIIIth Congr.Int.Primatol.Soc., Adelaide. (in press). (2001)

Suzuki, J.、Ishida, T.、Ohkura, S.、Udono, T.、Hayasaka, I. 和 Araithamkul, V.：“类人猿和小猿类的血浆 IGF-1 水平。”Proc.XVIIIth Congr.Int

Suzuki, J., Ohkura, S., Hayakawa, S.and Hamada, Y.: "Time series analysis of plasma insulin-like growth factor-I and gonadal steroids in adolescent Japanese macaques (Macacafuscata)."Journal of Reproduction and Development. 46. 157-166 (2000)

Suzuki, J.、Ohkura, S.、Hayakawa, S. 和 Hamada, Y.：“日本青少年猕猴 (Macacafuscata) 血浆胰岛素样生长因子-I 和性腺类固醇的时间序列分析。”生殖与发育杂志

Pranomo, Z. A. D. et al.: "A novel cryptic exon in intron 2 of the human dystrophin"Biochem. Biophys. Res. Commun.. 267. 321-328 (2000)

Pranomo, Z. A. D. 等人：“人类肌营养不良蛋白内含子 2 中的新型神秘外显子”Biochem。

Shimizu,Y., et al.: "Sero-and Molecular Typing of Duffy Blood Group in Southeast Asians and Oceanians."Human Biology. 72(3). 511-518 (2000)

Shimizu,Y., et al.：“东南亚和大洋洲人达菲血型的血清和分子分型。”人类生物学。

Suzuki,J.and Ishida,T.: "Age changes in plasma IGF-1 concentration in free-ranging Japanese macagues (Macaca fuscata)"Journal of Medical Primatology. (in press). 1-4 (2001)

Suzuki, J. 和 Ishida, T.：“自由放养的日本猕猴（Macaca fuscata）血浆 IGF-1 浓度的年龄变化”医学灵长类动物学杂志。