Editor's Statement This paper describes a quick method for quantitation of cell number in microcultures. Such procedures are valuable for the many situations in which minimizing cells and medium volume is desirable, although somewhat specialized equipment is required for the procedure. An alternative procedure for quantitation of cells in microtiter culture appeared previously in this journal (McCaffrey, et al., 24∶247–252).
Summary
A new method has been developed to count cells “in situ”, based on a fluorogenic enzyme assay that measures the activity of alkaline phosphatase. Increasing cell number was shown to correlate closely with alkaline phosphatase activity and this relationship did not change with time in culture. The alkaline phosphatase assay (ALP assay) was able to estimate relative cell numbers over a range from about 104 to 5×105 for many cell types, including Hep-2, a derivative of HeLa, several human colorectal cell lines SW1222, SW837, LS174T and HT29, a normal human diploid cell strain MRC5 and a rodent line NIH-3T3. The ALP assay is rapid and efficient, making it a useful method for studying growth assays.
Article PDF
Avoid common mistakes on your manuscript.
References
Freshney, R. I., ed. In: Culture of Animal Cells. New York: Alan Liss Inc.; 1984:199–204.
Carmichael, J.; Degraff, W. G.; Gazdar, A. F., et al. Evaluation of a Tetrazolium-based semiautomated colorimetric assay: Assessment ofchemosensitivity testing. Can. Res. 47:936–943; 1987.
Ishikawa, E.; Kato, K. Ultrasensitive enzyme immunoassay. Scand. J. Immunol. 8:43–55; 1978.
Wray, L. K.; Harris, H. Monoclonal antibodies against placental-like and intestinal-like alkaline phosphatases in a malignant human cell line. Biochem. 139:503–508; 1984.
McComb, R. B.; Bowers, G. N.; Posen, S., eds. In: Alkaline Phosphatase. New York: Plenum Press; 1979;51–112.
Stigbrand, T.; Millan, J. L.; Fishman, W. H. The genetic basis of alklaine phosphatase isoenzyme expression in isozymes: Current topics in biological and medical research ed. New York: Alan Liss Inc.; 6:93–117; 1982.
Jacobs, J. P.; Jones, C. M.; Baillie, J. P. Characteristics of a human diploid cell strain, designated MRC5. Nature 227:168–170; 1970.
Fogh, M.; Trempe, G. New tumour cell lines. In: Fogh, J., ed. Human tumour cellsin vitro. New York: Plenum Press; 1875:115–141.
Tom, B. H.; Rutzky, L. D.; Jakstys, M. M., et al. Human colorectal adeno carcinoma cells. 1. Establishment and description of a new line. In Vitro 12:180–191; 1976.
Leibovitz, A.; Stinson, J. C.; McCombs, W. B., III., et al. Classification of a human colorectal adeno carcinoma cell lines. Can. Res. 36:4562–4569; 1976.
House, W.; Waddell, A. Detection of mycoplasma in cell cultures. J. Pathol. Bacteriol. 93:125–132; 1967.
Bivic Le, A.; Arsanto, J.-P. Differential expression of alkaline phosphatase and ATPase activities in human colon carcinoma cell line HT-29. 18 during differentiation. Biology of the Cell 60:41–48; 1987.
Pignatelli, M.; Bodmer, W. F. Genetics and biochemistry of collagen binding triggered glandular differentiation in a human colon carcinoma cell line. Proc. Natl. Acad. Sci. USA 85:5561–5565.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Huschtscha, L.I., Lucibello, F.C. & Bodmer, W.F. A rapid micro method for counting cells “in situ” using a fluorogenic alkaline phosphatase enzyme assay. In Vitro Cell Dev Biol 25, 105–108 (1989). https://doi.org/10.1007/BF02624419
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02624419