培養膠質細胞の細胞化学

Cytochemistry of cultured glial cells

緒言

　培養細胞は生体内におけるものと同一ではないが，一般に個々の細胞が分離された状態にあること，さらにその細胞がin situの時に比して伸展されて存在する^1）点で（図1），細胞化学的検索，特に諸種物質の細胞内局在性の追求には好適の材料である。

　一方，培養細胞とくに培養した神経組織由来の細胞は，通常その周囲にあるplasma clotが，諸種薬剤の滲透を困難にするためか，その染色は一般に普通の切片標本に比して困難であつて，固定法，染色液の濃度，染色時間等について種々のmodificationを必要とする事が多い^2）3）。そのためか細胞化学的検索に対して前述したような利点があるにもかかわらず，培養細胞の細胞化学的研究は比較的少なく，Walker等の紫外線顕微鏡を用いてのデオキシリボ核酸（DNA）の研究^4），Kaufmann等の琥珀酸脱水素酵素の研究^5），Chèvremont等のアルカリ性フオスファターゼの研究^6），また，栽が国では近時，嶋井^7）の培養神経組織に関する研究等があるのみである^＊。

　1. The intracytoplasrmic granules in astrocytes grown in culures of cerebellar cortices obtained from puppies or kitten were cytochemically studied by means of various kinds of staining technics such its supravital staining with Janus green, Methylene blue, Neutral red, Toluidin blue, Azur B, and Sudan black as well as staining after fixation with Alcian blue, Sudan IV, Schiff's reagent Sudan black, PAS, Feulegn reaction, PFAS), Osmic acid Kolachev), Laxol fast blue, Schultz's method, Naphtol yellow s, Mallory's method. Kull's method and Jacobson's method.

　As a result of serial stai mg proced ores tested, it was concluded that the cytoplasmic granules which take Neutral red supravitally (NRG) are phospholipretein complex containing occasionally cholesterol and its esters.

　2. Furthermore the staining technics for the demonstration of enzymes were adapted and it was found that NRG contain acid phophoatase(Gomori's method), lipase (Tween 80) and a part of alkaline phosphataseiGomori's method), but Succinic, lactic, glutamic and malic dehydrogenase systems, adonosine triphosphotase, monoamine oxidase were not positive. It is duly postulated that NRG correspond in fact to "lysosome" described by de Duke et al. (1955).

　Lysosomes (NRG) were markedly decreased in number in fibroblasts cultured under anoxia attained by a modification of Schoettensack's method using alkaline pyrogallol. This decrease in numder of lysosome was accompanied by a concomitant appearance or increase in number of degenerative phospholipid granules devoid of protein moiety.

In addition to these findings, electron microscopic observation on cultured fibroblasts revealed dense bodies as well as bodies presumed to be lysosomes (Novikoff et al. : )