低カルシウム培養液を用いた新しい網膜色素上皮細胞の培養方法の確立

低钙培养基培养视网膜色素上皮细胞新方法的建立

• 批准号: 05771415
• 负责人: 宮良 長治
• 金额: $ 0.58万
• 依托单位: University of the Ryukyus
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05771415/
• 关键词: 網膜色素上皮細胞; カルシウム濃度; 牛胎児血清

1)牛胎児血清の濃度に関する結果。Medium199に1%bovine retinal extracts,insulin(10mug/ml),transferrin(5mug/ml),hydrocortisone(20nM),triiodothyronin(1nM),putrescine(0.3mug/ml),linoleic acid(0.3muM;complexed to bovine serum albumin),bovine serum albumin(10mug/ml)を加えると1%fetal bovine serumで培養可能であった。しかし、ヒト網膜色素上皮細胞では十分生育したが、ニワトリ網膜色素上皮細胞では増殖は弱く、なかなかconfluentにならなかった。2)低カルシウム下での網膜色素上皮細胞の培養結果上記の培養液ではヒト網膜色素上皮細胞は0.1mMのカルシウム下では培養可能で、細胞は増殖してconfluentになると浮遊してきた。この浮遊してきた細胞を別の培養皿に移すと、しばらくは培養皿に付着して増殖しconfluentになるとまた浮遊してきた。つまりトリプシンなどを使わなくても継代が可能であった。0.9mMでは細胞は培養皿に付着して生育し、浮遊しなかった。ニワトリ胚網膜色素上皮細胞は0.1mMの低カルシウム下では増殖不可能で、1.8mMとかなりの濃度を必要とした。以上よりヒト網膜色素上皮細胞は0.1mMカルシウム下で、無血清培養は不可能であったが1%にまで牛胎児血清の濃度を落とすことができ、トリプシンを使用なくても継代できた。ニワトリ胚網膜色素上皮細胞は同条件では不可能で、網膜色素上皮細胞の培養に関しては種差が存在することが明らかとなった。

1) the results of serum concentration test of bovine fetus. Medium199 complex 1%bovine retinal extracts,insulin (10mug/ml), transferrin (5mug/ml), hydrocortisone (20nM), triiodothyronin (1nM), putrescine (0.3mug/ml), linoleic acid (0.3mmm complexed to bovine serum albumin), bovine serum albumin (10mug/ml) may be difficult to detect. The pigmented epithelium cells of the reticular pigment epithelium of the mother and the mother were very pregnant, the proliferation of the pigment epithelial cells of the reticular pigment epithelium was weak, and the confluent cells were very weak. 2) the results of cell culture of membrane pigment epithelium in low temperature culture showed that the culture solution was used to detect the possibility of 0.1mM cell culture, confluent cell culture, cell culture and cell culture. There is no difference between the floating confluent and the petri dish, which means that the cells are not affected. I don't know. I don't know. I don't know. The 0.9mM cell culture dish pays for the birth and the buoyancy. It is impossible to colonize under the low temperature of the embryo membrane pigment epithelium (0.1mM), and it is necessary to colonize under the low temperature of 1.8 mm. It is impossible to reduce the serum concentration of bovine fetus by serum-free culture, and it is impossible to reduce the serum concentration of 1% fetal bovine fetus. The serum concentration of fetal bovine fetus is different from that of normal bovine fetus, and the serum concentration of fetal bovine fetus is low. It is not possible for the embryo membrane pigment epithelium cells to be cultured in the same condition, and there is a significant difference in the difference between the two groups.