Protein-free cell culture on polyvinyl formal surface.
Explore the source record for details and available documents.
Animals↗
Biomedical subjects
H Rabenau
Publications and source records attributed to H Rabenau.
Polyvinyl formal surface promotes continuous growth of Vero cells in protein-free medium.
The effect of polyvinyl formal (PVF) culture surface on the growth of 10 mammalian continuous cell lines, including Swiss 3T6, NCTC clone 929 L, BHK-21 clone 13, CHO-K1, PK 15, A 431, HeLa, MDCK, LLC-MK2 and Vero in protein-free 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 supplemented with trace elements and L-ascorbic acid 2-phosphate, was investigated. Most of the cell lines showed only some initial proliferation on PVF similar to the polystyrene (PS) surface of commercially available culture flasks. In contrast, proliferation of monkey kidney cell line Vero was by far greater on PVF than on PS or poly-D-lysine treated culture surface. In addition, Vero cells on PVF could be subcultured in the protein-free medium without any significant decrease of growth rate in successive passages. These results showed that PVF provides a culture surface which selectively promotes continuous growth of Vero cells in protein-free, chemically defined medium.
Animals↗
HeLa cells grown continuously in protein-free medium: a novel model for the study of virus replication.
Human carcinoma of the cervix cell line HeLa, adapted to continuous growth in a protein-free chemically defined medium, was used as substrate for the replication of several human pathogenic viruses. Growth characteristics of the cells designated as HeLa-PF in protein-free 1:1 nutrient mixture of Dulbecco's modified MEM and Ham's F-12 supplemented with L-ascorbic acid 2-phosphate were similar to those of the cells grown in a serum-supplemented medium. After 30 months (135 subcultures) in the protein-free medium, HeLa-PF cells were infected with poliovirus types 1, 2 and 3; adenovirus types 2 and 5 and herpes simplex virus type 1. Both adenoviruses and polioviruses developed in HeLa-PF cells titers and showed cytopathic effects comparable to those obtained in conventionally grown and maintained cells; in contrast, significantly lower herpes simplex virus type 1 titers and changed characteristics of the cytopathic effects were observed in HeLa-PF cells. The results show that HeLa-PF cells grown continuously in protein-free medium provide a unique system for the study of virus replication.
Adenoviridae↗
Effect of aciclovir on the replication of herpes simplex virus type 1 in MA-104 cell line resistant to aciclovir.
A culture of monkey kidney cells (MA-104) resistant to aciclovir (CAS 59277-89-3) was established. The resistant cells were continuously grown in medium containing 200 mumol/l aciclovir whereas normal cells died in such medium. The doses of aciclovir required for 50 and 90% reduction in virus yield were for several Herpes simplex virus Type 1 clinical isolates up to 5-fold higher in resistant than in normal MA-104 cells; in contrast, doses required for 99% reduction in virus yield could be as much as 50-fold higher in the resistant cells. Virus yields developed in the presence of 30 mumol aciclovir in culture medium were at least 10(3)-fold higher in resistant than in normal MA-104 cells. The results showed that the antiviral effect of aciclovir may be significantly decreased in the cell line which developed resistance to aciclovir.
Acyclovir↗
Quality control testing of surfaces for mammalian cell culture using cells propagated in a protein-free medium.
Mouse NCTC clone 929 L (L-929) cells were propagated continuously for 3 years as monolayers in a protein-free chemically-defined medium. These cells, designated L-929-WS, were used for quality control testing of the surfaces of commercially available cell culture plastic flasks. Differences in attachment and saturation density of L-929-WS cells in a protein-free culture medium were taken to define various levels of quality of the culture vessels tested. The rate of attachment and growth of L-929-WS cells on a surface of a given quality correlated directly with that of human embryonal fibroblasts and embryonal epithelial cells grown in a serum-free medium supplemented with growth factors and hormones. L-929-WS cells propagated continuously in a protein-free medium provide a simple and sensitive assay system for more general quality control testing of surfaces used for the culture of monolayer cells.
Animals↗
[Atypical pathogen spectrum in community-acquired infections of the lower respiratory tract in childhood].
Over a period of 14 months, we obtained nasal and throat swabs for virological examinations from patients, admitted to hospital with suspected pneumonia. Fifty-five cases of perihilar bronchitis and 103 cases of pneumonia were diagnosed based on radiological criteria. In 36% (n = 57) of the children, a pathogen could be determined by antigen ELISA. We found the following spectrum: Respiratory syncytial virus: 42%; Influenza A: 8%; Influenza B: 10%; Parainfluenza 1: 3%; Parainfluenza 2: 10%; Parainfluenza 3: 2%; Chlamydia: 17%; and Mycoplasma pneumoniae (serology only): 7%. In a control group of 59 healthy children, only one child showed a pathogen. In the past, viruses and Chlamydia have been underestimated as causes of lower respiratory tract infections. Antigen determination by ELISA has proved to be a reliable and rapid method for their detection.
Adolescent↗
A novel assay system for quality control testing of surfaces for mammalian cell culture.
Explore the source record for details and available documents.
Animals↗
Congenital rubella syndrome after maternal reinfection.
This report concerns a case of congenital rubella syndrome (CRS) with atypical immune response affecting an infant whose mother had repeated evidence of immunity before pregnancy. Laboratory diagnosis of CRS could only clearly be achieved by virus isolation after the second month of life despite typical clinical features of CRS and multiple organ involvement. After the first month of age, low concentrations of specific IgM antibodies were revealed by ELISA and confirmed by a reference test system (IgM-specific haemagglutination inhibition assay). Persistent and increasing high levels of IgM antibodies were detected only after the 6th month of life. Later on IgG antibody levels decreased. Immunological investigations showed an IgG1-hypoglobulinaemia. The unusual feature of the present case report is not only the failure of the maternal rubella immunity to prevent CRS, but the defect of the child's immune system, probably attributable to congenital infection. As a consequence, laboratory diagnosis of CRS could not be achieved initially by the proved serological methods.
Adult↗
Replication of human herpesvirus type 6 (strain AJ) in JJHan cells grown in protein-free medium.
The human T-cell line JJHan, commonly used as a substrate for human herpesvirus type 6 (HHV-6) replication, was established in a protein-free 1:1 mixture of Ham's F12 and Iscove's modified Dulbecco's medium. After 250 passages (4 years) in protein-free medium, the cells, designated JJHan-PF, were cocultured with JJHan cells chronically infected with strain AJ of HHV-6 conventionally grown in serum-supplemented medium. The production of viral antigen was monitored by indirect immunofluorescent antibody staining. The infectivity of the virus in JJHan-PF cells was similar to that in JJHan cultures as shown by HHV-6 antigen detection using a convalescent phase serum. The results show that JJHan-PF cells provide a sensitive substrate for HHV-6 replication without the interference of serum or other proteins usually added to the culture medium.
Cell Line↗
Cytotoxicity of L-cycloserine against human neuroblastoma and medulloblastoma cells is associated with the suppression of ganglioside expression.
BACKGROUND: Human neuroblastoma and medulloblastoma express abnormal ganglioside patterns especially GD2 and GM2 which are important for tumour growth. We tested the effects of L-cycloserine (L-CS), a potent inhibitor of synthesis of glycosphingolipids, on the growth, viability and expression of GD2 and GM2 in neuroblastoma and medulloblastoma cells. MATERIALS AND METHODS: The cytotoxic effects of L-CS were tested using the MTT dye reduction assay on four neuroblastoma (IMR-32, SK-N-SH, UKF-NB-2 and UKF-NB-3), two medulloblastoma (D283 and D341) and normal human fibroblasts and epithelial cell lines. In some experiments cytotoxicity of L-CS was tested in the presence of exogenous GD2 and GM2. The expression of GD2 and GM2 was analysed by flow cytometry. The antitumoral effects of L-CS in vivo were assessed on established xenografts of UKF-NB-3 or D283 cells in athymic (nude) mice using systemic administration of the drug (150 mg/kg intraperitoneally, once per day on 20 consecutive days). RESULTS: In vitro experiments revealed that L-CS was toxic for tumour cells at concentrations ranging from 0.5 to 20 micrograms/ml without any significant effects on normal fibroblasts and epithelial cells. L-CS treatment of UKF-NB-3 and D283 cells significantly inhibited expression of GD2 and GM2. The addition of exogenous GD2 and GM2 to culture medium partially prevented cytotoxic effects of L-CS on tumour cells. In vivo treatment resulted in complete tumour regression of UKF-NB-3 xenografts whereas growth of D283 xenografts was reduced by 60%. CONCLUSIONS: L-CS is a selective antitumoral agent for neuroblastoma and medulloblastoma cells with the ability to reduce expression of tumour associated gangliosides. In vivo experiments suggest that L-CS may be effective drug for treatment of neuroblastoma and medulloblastoma.
Animals↗