동결건조법에 있어 Nocardia mediterranei의 세포막 손상과 재수화 방법에 따른 생존도

Membrane Injury of Nocardia mediterranei upon Lyophilization and Viability Depending on Rehydration Methods

동결건조법이 Nocardia mediterranei의 세포막 손상에 미치는 영향을 3H-thymidine 표지에 의한 DNA 유출 방법과 전자현미경으로 조사하였으며, 재수화 과정이 생존도에 미치는 영향을 연구하였다. 그 결과 N.mediterranei의 동결건조시 세포벽과 세포막의 손상이 세포 사멸의 원인으로 판명되었다.

In order to examine the viability depending on rehydration process and membrane injury of Nocardia mediterranei upon lyophilization, We labeled $3^H$-thymidine in deoxyribonucleic acid of N. mediterrranei to obtain information on the mechanisms of injury caused by lyophilization. Suspensions of rehydrated cells were incubated with added DNase in a buffer solution. Extracellular radioactivity levels appeared to be high in the rehydrated solutions after lyophilization than freezing-thawing. Thus, the membrane systems were injured by lyophilization, but not ovenvhelmed. These considerations were confirmed by electron microscopy. In effects of rehydration, the cell membrane was seriously damaged by strong atmospheric pressure as soon as the inner ampule was opened, but this was not the case without admitting air under vacuum. N. rnediterranei cells, with no additives, were lyophilized and reconstituted without admitting air, virtually about 84% of the cells were viable.