Fundamental study on establishment and application of embryostem cell(ESC) in domestic animals.

家畜胚胎干细胞（ESC）的建立及应用基础研究。

基本信息

批准号：
06454122
负责人：
UMEZU Motoaki
金额：
$ 4.67万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

Embryo stem cell (ESC) is the cell line originated from embryo and maintatined in undifferentiated state with pluripotency or totipotency. The cell is utilized in model of gene disease and therapy and establishment of cell lines is not fully successful in another animals except in mouse. In 1995-1996, establishment of ESC lines was tried from embryo originated cell in cow and goat as a domestic animal and in rat as an experimental animal.In cow, oocytes harvested from ovaries of abattoir were subjected to in vitro maturation, fertilization and development to blastocysts and the inner cell mass (ICM) from blastocysts were tried to passage culture. The tertiary passage was possible when mouse embryo fibro-blast cell (MEF) or STO cell was used as feeder cell. Sampling of bovine ovaries was stopped because the slaughter house in Sendai-shi was closed for outsider caused of 0-157 affair.In goat, morula, ICM and embryo just prior to hatching that were originated from in vivo fertilized ova were subjected to passage culture using MEF cell or STO cell as feeder cell and undifferentiated state was maintained in secondary culture using ICM.In rat, in vivo sampled molula, ICM and primordial germ cell (PCM) of 9 dpc were subjected to passage culture and the effect of feeder cell was screened using MEF,rat embryo fibro-blast (REF), and S1^4-m220. Colony formation with undifferentiation was confirmed in ICM and PGC when REF or S1^4-m220 was used as feeder. PGC was passaged for fifth culture and further examination is suggested to need the cell line formation from PGC.

胚胎干细胞（ESC）是起源于胚胎、维持未分化状态、具有多能性或全能性的细胞系。该细胞用于基因疾病和治疗的模型，并且细胞系的建立在除小鼠之外的其他动物中并未完全成功。 1995-1996年，以牛和山羊为家畜，以大鼠为实验动物，尝试建立ESC系。在牛中，从屠宰场卵巢收获的卵母细胞进行体外成熟、受精和发育。囊胚和来自囊胚的内细胞团（ICM）尝试进行传代培养。当小鼠胚胎成纤维细胞（MEF）或STO细胞用作饲养细胞时，第三次传代是可能的。由于0-157事件导致仙台市屠宰场对外人关闭，牛卵巢取样被停止。在山羊、桑椹胚、ICM和即将孵化的胚胎中，来自体内受精卵的传代培养使用MEF细胞或STO细胞作为饲养细胞，并使用ICM在继代培养中维持未分化状态。在大鼠体内，采集软体、ICM和原始胚芽对9 dpc细胞(PCM)进行传代培养，并使用MEF、大鼠胚胎成纤维细胞(REF)和S1^4-m220筛选饲养细胞的效果。当使用 REF 或 S1^4-m220 作为饲养层时，在 ICM 和 PGC 中证实了未分化的集落形成。 PGC 传代进行第五次培养，建议进一步检查是否需要从 PGC 形成细胞系。