| Applications |
The cells can be used in adherence assays as a target cell for malaria infected erythrocytes. Functional expression of the transfected receptor molecules on CHO-ICAM-1 was confirmed using parasites with different cytoadherence phenotypes and cytoadherence inhibition experiments with a panel of anti ICAM-1 antibodies. The use of isolates obtained from malaria patients in The Gambia confirmed the applicability of using these cells in assays for laboratory studies. |
| Derivation |
The CHO-ICAM-1 cell line was constructed at DNAX Research Institute, Palo Alto, California by transfecting Chinese hamster ovary (CHO) cells with linearized cDNA of human ICAM-1 in pCDM8 by electroporation in a 10 fold molar excess over pBJ1-Neo. The use of isolates obtained from malaria patients in The Gambia confirmed the applicability of using these cells in assays for laboratory studies. |
| Comments |
The CHO-ICAM-1 cell line was constructed at DNAX Research Institute, Palo Alto, California by transfecting Chinese hamster ovary (CHO) cells with linearized cDNA of human ICAM-1 in pCDM8 by electroporation in a 10 fold molar excess over pBJ1-Neo. Stable transfectants were then selected by growth in culture medium containing G418. CHO-ICAM-1 cells express high levels of human ICAM-1 (a ligand for erythrocytes infected with the human malaria parasite, Plasmodium falciparum). The cells can be used in adherence assays as a target cell for malaria infected erythrocytes. Functional expression of the transfected receptor molecules on CHO-ICAM-1 was confirmed using parasites with different cytoadherence phenotypes and cytoadherence inhibition experiments with a panel of anti ICAM-1 antibodies. The use of isolates obtained from malaria patients in The Gambia confirmed the applicability of using these cells in assays for laboratory studies. A culture submitted to the ATCC in August 1993 was found to be contaminated with mycoplasma, and the line was cured by a 21 day treatment with BM Cycline. |
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:6 to 1:10
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| References |
Hasler T, et al. An improved microassay for Plasmodium falciparum cytoadherence using stable transformants of Chinese hamster ovary cells expressing CD36 or intercellular adhesion molecule-1. Am. J. Trop. Med. Hyg. 48: 332-347, 1993. PubMed: 7682381
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.
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