現在、低酸素の条件で培養を行うためには、窒素ガスボンベを使用する。我々は窒素ガスボンベに代わる窒素発生装置を作製し、それを炭酸ガスフラン器に接続し、低酸素培養条件を作ることに成功した。この窒素発生装置は、空気を分子炭を充填したカラムに通すことにより窒素ガスのみを精製、濃縮できるもの篩である。この装置を用いた1%、5%、10%、15%酸素条件における種々の細胞の増殖能の検討を行った。その結果、この低酸素条件下でコロニー形成率および増殖率が上昇する細胞系が多かった。

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本研究では当科で作製したEGF受容体(EGF-r)に対するモノクローナル抗体(MoAb)12-93抗体のヌードマウス移植ロ腔癌の増殖に及ぼす影響さらに、12-93抗体結合LAK細胞を用いた口腔癌に対する特異的癌養子免疫療法の検討を行った。
12-93抗体はヌードマウス移植口腔扁平上皮癌細胞および唾液腺腺癌の増殖を抑制した。12-93抗体結合LAK細胞は、EGF-rを過剰発現している口腔癌に対しては抗体非結合LAK細胞と比較して高い細胞障害活性を示した。一方、EGF-rを発現していないRaji細胞に対しては抗体非結合LAK細胞と同等の活性を示した。
これらの結果は、EGF-rに対するMoAb 12-93抗体は抗体単独で抗腫瘍活性を有し腫瘍特異抗体に代わる有用な抗体であること、さらに12-93抗体結合LAK細胞は特異的癌養子免疫療法を行う上で効果的な方法であることを示唆している。

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Increase in the incubation temperature of infected Aedes albopictus clone C6/36cells from its optimal growth temperature of 28°C to 32 and 37°C showed a concomitant rise in the production of dengue virus type 2 antigen and genomic RNA. The quantitative reverse transcription-polymerase chain reaction using an internal standard as a measuring tool for the viral genomic RNA showed that RNA production was higher in the cells and in the extracellular fluid from the cultures incubated at 32 and 37°C than at 28°C. The presence of viral antigen and viral genomic RNA was also detected 1-2days earlier in the cultures incubated at elevated temperatures.

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Combined effects of methyl methanesulfonate (MMS) and ethyl methanesulfonate (EMS), which were administered successively for each 3 h at different times, or with an interval time of incubation in normal medium for 3 h between treatments with two chemicals were examined on survival and induction of 6TG-resistant and OUA-resistant mutations in Chinese hamster V79 cells.
Pre-treatment with EMS decreased markedly the survival of cells treated with MMS compared with that of cells treated simultaneously with both chemicals. The insertion of interval time between treatments with both chemicals restored the cell survival to that treated with EMS alone. The pre-treatment with MMS also decreased the survival of cells treated with EMS more markedly than that of cells treated with two chemicals simultaneously.
When cells were pre-treated with EMS, the frequency of 6TG-resistant mutations induced by MMS enhanced to be higher than that of cells treated with both chemicals simultaneously. In this case, the restoration effect of incubation time between treatments with two chemicals was found. On the other hand, pre-treatment with MMS, no detectable change in the frequency of 6TG-resistant mutations induced by EMS was found, compared with that in cells treated with both chemicals simultaneously.
In the induction of OUA-resistant mutations induced by EMS, pre-treatment with MMS increased the mutation frequency compared with that in cells treated with both chemicals simultaneously. Also in the simultaneous treatment with both chemicals, the frequency of QUA-resistant mutations was higher than that of mutations induced by EMS alone. The insertion of interval between two treatments with both chemicals reduced the mutation frequency.

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