A novel assay system for quanlity control testing of surfaces for mammalian cell culture

CinatlJr., Jindrich; Činátl, Jaroslav; Rabenau, Holger F.; Türk, Marina; Ahn, Junho; Doerr, H. W.

doi:10.1007/bf02624606

Cited by 4 publications

(2 citation statements)

References 7 publications

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“…Neverthe less, such cells have not had general applica tion. It has been repeatedly suggested that suc cessful growth of the cells in a protein-free medium depends on several factors including high quality chemicals, pure water and chemi cally clean glass or plastic ware [14,[22][23][24]. It is possible that the quality of these factors was difficult to obtain in the past and, thus, the culture technique could only be used in a few laboratories.…”

Section: Discussionmentioning

confidence: 99%

HeLa Cells Grown Continuously in Protein-Free Medium: A Novel Model for the Study of Virus Replication

Cinatl¹,

Rabenau²,

Kornhuber

et al. 1992

Intervirology

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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.

show abstract

Section: Discussionmentioning

confidence: 99%

HeLa Cells Grown Continuously in Protein-Free Medium: A Novel Model for the Study of Virus Replication

Cinatl¹,

Rabenau²,

Kornhuber

et al. 1992

Intervirology

View full text Add to dashboard Cite

show abstract

“…Despite the possible practical advan tages of such cells for different purposes including virus production, this culture technique has not found general application. It has been repeatedly suggested that success ful cell growth in a protein-free medium depends on sev eral factors including high-quality chemicals, pure water and chemically clean glass or plastic ware [22][23][24][25], It is possible that the quality of these factors was difficult to obtain in the past, and thus the culture technique could only be used in a few laboratories. In the present study, we observed effects of different lots of protein-free DMEM-F12 medium obtained from several suppliers on the growth of HeLa-PF cells in suspension culture.…”

Section: Discussionmentioning

confidence: 99%

Suspension Culture of HeLa Cells in Protein-Free Medium: Sensitivity to Human Pathogenic Viruses

Činátl

Wichelhaus²,

Weber³

et al. 1994

Intervirology

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Human adherent HeLa-PF cells grown for 5 years in a protein-free 1:1 nutrient mixture of Dulbecco’s modified MEM and Ham’s F12 (DMEM-F12) were established in suspension culture. The cells grew in protein-free DMEM-F12 (using magnetically stirred flasks) as a monodisperse suspension with a population doubling time of 28 h. The cells were infected with poliovirus types 2 and 3, herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), respiratory syncytial virus (RSV), echovirus 6 and adenoviruses 3 and 7. Polioviruses replicated in suspension culture of HeLa-PF cells to a similar extent as in protein-free and serum-supplemented adherent cultures. RSV and HSV developed significantly lower titers while adenoviruses and echovirus 6 developed significantly higher titers in suspension than in adherent cultures. The results show that suspension culture of HeLa-PF may both provide the advantage of a high virus yield and enable cultivation of viruses which are not contaminated by serum or other proteins usually added to the culture medium.

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Role of serum vitronectin and fibronectin in adhesion of fibroblasts following seeding onto tissue culture polystyrene

Steele¹,

Johnson²,

Underwood³

1992

J. Biomed. Mater. Res.

172

130

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The suitability of polymeric biomaterials as surfaces for the attachment and growth of cells has often been investigated in tissue culture. In this study the contribution that adsorption of serum fibronectin (Fn) or vitronectin (Vn) make to the attachment and spreading of fibroblast cells during the first 90 min following seeding was determined for two modified tissue culture polystyrenes, as model biomaterial surfaces. The amount of serum Vn and Fn which adsorbed onto tissue culture grade polystyrene (TCP) from different serum concentrations over the range of 0.1-30% (v/v) were determined and compared to attachment of cells of the BHK-21 and HT1080 fibroblast lines. There was no simple correlation between the amount of Fn or the amount of Vn adsorbed and cell attachment and spreading. The requirement for Fn or Vn for attachment and spreading of BHK-21 or HT1080 cells onto modified polystyrene (either TCP or to Primaria) during the first 90 min of cell culture was directly tested by selective removal of Fn or Vn from the serum prior to addition to the culture medium. Attachment and spreading of BHK-21 or HT1080 cells onto TCP or Primaria surfaces were reduced in a concentration-dependent manner when the cells were seeded in medium containing 2% (v/v) or higher concentrations of Vn-depleted serum. BHK-21 cells or HT1080 cells seeded in medium containing Fn-depleted serum (which contained Vn) attached and spread onto TCP or Primaria. Both BHK-21 cells and HT1080 cells failed to attach to TCP or Primaria when seeded in medium containing serum depleted of both Vn and Fn. The requirement for serum Vn or Fn for fibroblast attachment to TCP was also tested using cells of a human dermal fibroblast strain. The attachment of the dermal fibroblasts to TCP during the first 90 min of culture was not decreased by depletion of Vn from the 15% (v/v) serum, but there was a reduction in the proportion of the attached cells which had spread. Selective depletion of serum Fn did not have any effect on either cell attachment or spreading. Our results show that for fibroblast cells, particularly with cell lines such as BHK-21 or HT1080 but also with cell strains, the first binding of cells onto tissue culture polystyrene when plated in medium containing serum is a result of adsorption onto the surface of serum Vn. The adsorption of serum Vn onto the surface overcomes the effect of serum components which tend to decrease cell attachment.

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A novel assay system for quanlity control testing of surfaces for mammalian cell culture

Cited by 4 publications

References 7 publications

HeLa Cells Grown Continuously in Protein-Free Medium: A Novel Model for the Study of Virus Replication

HeLa Cells Grown Continuously in Protein-Free Medium: A Novel Model for the Study of Virus Replication

Suspension Culture of HeLa Cells in Protein-Free Medium: Sensitivity to Human Pathogenic Viruses

Role of serum vitronectin and fibronectin in adhesion of fibroblasts following seeding onto tissue culture polystyrene

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