単一細胞に分離した人間胎児の組織再形成(予報)

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This paper describes briefly the methods and results of a rotation-mediated cultivation of dissociated human embryonic cells. The whole body of a two-month male human foetus was cut into shreds with scissors. The shreds were suspended in a 0.2% trypsin solution, approximately 10 times in volume, and stirred at 37℃ for 30 minutes. Then an equal amount of culture medium was added to the tissue suspension thus obtained. The tissue was dispersed into single cells by flushing them briskly, use being made of a 20ml syringe fitted with a needle (1.5mm inner diameter). After having been centrifuged at 3000rpm for 15 minutes, the sediments were washed with three changes of the culture medium, and finally re-suspended in the medium, with adjustment to 10^7 cells per ml. The culture medium used for cultivation was prepared after Horikawa's formula (1961). One and a half ml of the cell suspension thus prepared were transferred to 10ml Erlenmeyer flasks in each. The flasks were horizontally rotated on a platform of a gyratory shaker with a 3/4inch eccentric rotary motion at a constant speed of 60 rpm at 37℃. After 1 to 2 hours of cultivation, numerous small clusters of cells appeared in flask cultures. With the lapse of time those clusters increased in size by accretion of free cells. By 24 hours, about 50 rounded or oblong aggregates of cells with bumpy surface, 0.1 to 1.0mm in diameter being 0.3mm on the average, were present in each flask. In addition, about 5×10^6 cells per ml always remained discreted being free in the medium. It was seen in paraffin sections through the cell-aggregates that there were certain histotypic groups of cells. Those groups were formed by a selective sorting out of cells according to their specific type. Among those groups of cells, a group of cartilagious cells forming hyaline cartilage was most remarkable and clearly identifiable in nature. Cells with large-sized vacuoles were dispersed in the interstitial substance. This tissue was sharply demarcated from the surrounding tissues by several layers of flattened cells. It is thus evident that the results of the present experiments with dissociated cells from a human foetus supplement similar features reported by Moscona (1961a, 1962) and by Steinberg (1962 b, c, d) in chick embryos.
社団法人日本動物学会の論文
1963-04-15

単一細胞に分離した人間胎児の組織再形成(予報)

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