RESUMO
The author describes a multiplate settling vessel in which it is possible to separate the cells from the growth medium even if in large scale (up to 1.800 1) in less than 24 hrs, discarding completely the supernatant.
Assuntos
Separação Celular/métodos , Técnicas Citológicas , Animais , Aphthovirus/imunologia , Centrifugação , Células Clonais , Cricetinae , Meios de Cultura , Rim , Vacinas Virais/isolamento & purificação , Cultura de Vírus/métodosRESUMO
From BHK 21 cl. 13 suspensions cells we have isolated a virus serologically related to the Minute Virus of Mice (MVM); the same virus has been isolated from the same cells in U.K. in 1980. Besides isolating this virus from large scale cultures, we have also isolated it from many of the sublines stored in nitrogen we received at different times from different laboratories. At this time it is difficult to ascertain if the contamination originated from the serum or from other components of the medium; furthermore we do not know if many of the cells of our stock became contaminated in our laboratory or if they were already contaminated.
Assuntos
Células Clonais/microbiologia , Vírus Miúdo do Camundongo/isolamento & purificação , Parvoviridae/isolamento & purificação , Animais , Cricetinae , Efeito Citopatogênico Viral , Rim , Vírus Miúdo do Camundongo/imunologia , Parvoviridae/imunologiaRESUMO
The problems related to the use of serum in cell culture are reviewed. The possibility of substituting the serum with peptones has already been shown. Different peptones have been tested: one of the best is a peptone obtained from meat and casein pepsin pancreatically digested. BHK 21/13 cells were cultivated in serum-free media for 35 passages. The 0.81 cultures without automatic pH control had a cycle length of 3 days; starting with concentrations of 0.4 X 10(6) cells/ml, concentrations higher than 3 X 10(6) cells/ml were often obtained. We did not obtain satisfactory results when the volume of cultivation was increased to more than 6 1, perhaps because of different requirements for agitation, pH, O2. It is also necessary to check whether the results obtained up to now are consistent with reference to the source of reagents and cell strain used. The absence of serum in the medium did not influence virus replication when infectivity and complement fixation titers were tested.