Skip to main content
Log in

From growth factor dependence to growth factor responsiveness: The genesis of an alveolar macrophage cell line

  • Published:
In Vitro Cellular & Developmental Biology Aims and scope Submit manuscript

Summary

A rat pulmonary alveolar macrophage (PAM) cell line (NR8383) was initiated in culture in the presence of a gerbil lung cell conditioned medium (GLCM), and has been propagated continuously for over 36 mo. When examined at different times throughout this in vitro period, NR8383 exhibited characteristics typical of macrophages: (a) Zymosan ingestion was seen in 90 to 98% of the cells examined; (b)Pseudomonas aeruginosa phagocytosis in 50 to 80%; (c) Nonspecific esterase activity in >95%. During the first 6 mo., the PAM replicated with doubling times approximating 15 to 20 d. Throughout this period, GLCM dependence was evident. After 27 wk in vitro, NR8383 replication increased markeldy, and within 2 wk, the doubling time was less than 48h. NR8383 was readily monitored by [3H]thymidine (TdR) blastogenesis assay. In the presence of GLCM uptake of [3H]TdR was fivefold greater than in control cultures. Adherence and growth kinetics were effectively controlled by modulation of GLCM or serum content in culture medium. It was demonstrated that PAM growth factor(s) is ubiquitous, not species-specific, and under certain conditions, may be derived from “endogenous” sources of persisting non-PAM populations within the parent, uncloned line NR8383. Cloned progeny remain devoid of non-PAM “feeder” cells, but retain macrophage properties, including interleukin-1 secretion, Fc receptors, and H2O2 production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Boyd, R. L.; Ramphal, R.; Rice, R., et al. Chronic colonization of rat airways withpseudomonas aeruginosa. Infect. Immun. 39:1403–1410; 1983.

    PubMed  CAS  Google Scholar 

  2. Bradley, T. R.; Metcalf, D. The growth of mouse bone marrow cellsin vitro. Aust. J. Exp. Biol. Med. Sci. 44:289–299; 1966.

    Google Scholar 

  3. Broxmeyer, H. F.; Ralph, P.In vitro regulation of a mouse myelomonocytic leukemia line adapted to culture. Cancer Res. 37:3578–3584; 1977.

    PubMed  CAS  Google Scholar 

  4. Bryne, P. V.; Guilbert, L. J.; Stanley, E. R. Distribution of cell bearing receptors for a colony-stimulating factor (CSF-1) in murine tissues. J. Cell Biol. 91:848–853; 1981.

    Article  Google Scholar 

  5. Cahoon, B. E.; Mills, J. Development and characterization of a continuous line of alveolar macrophages from C3H/HEJ mice. J. Reticuloendothel. Soc. 29:357–367; 1981.

    PubMed  CAS  Google Scholar 

  6. Chen, T. R.In situ detection of mycoplasma contamination in cell cultures by fluorescent Hoechst 33258 stain. Exp. Cell Res. 104:255–262; 1977.

    Article  PubMed  CAS  Google Scholar 

  7. Cohen, A. B.; Cline, M. J. The human alveolar macrophage: Isolation, cultivationin vitro and studies of morphologic and functional characterization. J. Clin. Invest. 50:1390–1398; 1971.

    Article  PubMed  CAS  Google Scholar 

  8. Golde, D. W.; Byers, L. A.; Finley, T. N. Proliferative capacity of human alveolar macrophage. Nature 247:373–375; 1974.

    Article  PubMed  CAS  Google Scholar 

  9. Hart, P. D.; Young, M. R. Interference with normal phagosomelysosome fusion in macrophages, using ingested yeast cells and suramin. Nature 256:47; 1975.

    Article  PubMed  CAS  Google Scholar 

  10. Hocking, W. G.; Golde, D. W. The pulmonary-alveolar macrophage. N. Engl. J. Med. 301:580–587; 1979.

    Article  PubMed  CAS  Google Scholar 

  11. Kaplan, D. R.; Henkel, T. J.; Braciale, V., et al. Mycoplasma infection of cell cultures: Thymidine incorporation of culture supernatants as a screening tests. J. Immunol. 132:9–11; 1984.

    PubMed  CAS  Google Scholar 

  12. Karnovsky, M. J. A formaldehyde-glutaraldehyde fixative of high osmolarity for use in electron microscopy. J. Cell Biol. 27:137a (abstract); 1965.

    Google Scholar 

  13. Li, C. Y.; Lam, K. W.; Yam, L. T. Esterases in human leukocytes. J. Histochem. Cytochem. 21:1–12; 1973.

    PubMed  CAS  Google Scholar 

  14. Lin, H.; Kuhn, C.; Kuo, T. Clonal growth of hamster free alveolar cells in soft agar. J. Exp. Med. 142:877–886; 1975.

    Article  PubMed  CAS  Google Scholar 

  15. Metcalf, D. The granulocyte-macrophage colony stimulating factors. Science 229:16–22; 1985.

    Article  PubMed  CAS  Google Scholar 

  16. Morahan, P. S. Macrophage nomenclature: Where are we going? (Editorial). J. Reticuloendothel. Soc. 27:223–231; 1980.

    PubMed  CAS  Google Scholar 

  17. Muschel, R. J.; Rosen, N.; Rosen, O. M. Modulation of Fc-mediated phagocytosis by cyclic-AMP and insulin in a macrophage-like cell line. J. Immunol. 119:1813–1820; 1977.

    PubMed  CAS  Google Scholar 

  18. Myrvik, Q. N.; Leake, E. S.; Fariss, B. Studies on pulmonary alveolar macrophages from the normal rabbit: A technique to procure them in a high state of purity. J. Immunol. 86:128–132; 1961.

    PubMed  CAS  Google Scholar 

  19. Naum, Y. Growth of pulmonary alveolar macrophagesin vitro: Responses to media conditioned by lung cell lines. Cytobios 14:211–220; 1975.

    Google Scholar 

  20. Pantazis, C. G.; Kniker, W. T. Assessment of blood leukocyte microbial killing by using a new fluorochrome microassay. J. Reticuloendothel. Soc. 26:155–170; 1979.

    PubMed  CAS  Google Scholar 

  21. Rabinovitch, M.; DeStefano, M. J. Macrophage spreadingin vitro I. Inducers of spreading. Exp. Cell Res. 77:323–324; 1973.

    Article  PubMed  CAS  Google Scholar 

  22. Ralph, P. Continuous cell lines with properties of mononuclear phagocytes. In: Adams, D. O.; Edelson, D. J.; Koren, H., eds. Methods for studying mononuclear phagocytes. New York: Academic Press; 1981:155–173.

    Google Scholar 

  23. Ruch, W.; Cooper, P. H.; Baggiolini, M. Assay of H2O2 production by macrophages with homovanillic acid and horseradish peroxidase. J. Immun. Methods 63:347–357; 1983.

    Article  CAS  Google Scholar 

  24. Senior, R. M.; Campbell, E. J.; Villiger, B. Obtaining and culturing human and animal alveolar macrophages. In: Adams, D. O.; Edelson, D. J.; Koren, H., eds. Methods for studying mononuclear phagocytes. New York: Academic Press; 1981:69–83.

    Google Scholar 

  25. Simon, P. L.; Laydon, J. T.; Lee, J. C. A modified assay for interleukin 1 (IL-1). J. Immunol. Methods 84:85–94; 1985.

    Article  PubMed  CAS  Google Scholar 

  26. Soderland, S. C.; Naum, Y. Growth of pulmonary alveolar macrophageshagesin vitro. Nature 245:150–152; 1973.

    Article  PubMed  CAS  Google Scholar 

  27. Stanley, E. R.; Guilbert, L. J. Methods for the purification, assay, characterization and target cell binding of a colony stimulating factor (CSF-1). J. Immunol. Methods 42:253–284; 1981.

    Article  PubMed  CAS  Google Scholar 

  28. Stewart, C. C.; Lin, H. Macrophage growth factor and its relationship to colony stimulating factor. J. Reticuloendothel. Soc. 23:269–285; 1978.

    PubMed  CAS  Google Scholar 

  29. Tooze, J. The molecular biology of tumor viruses. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory: 1979.

    Google Scholar 

  30. Virolaminen, M.; Defendi, V. Dependence of macrophage growthin vitro upon interaction with other cell types. In: Defendi, V.; Stoker, M., eds. Growth regulating substances for animal cells in culture. 1967:67–85.

  31. Warr, G. A.; Martin, R. R. Histochemical staining andin vitro spreading of human pulmonary alveolar macrophages: variability with cigarette smoking status. J. Reticuloendothel. Soc. 23:53; 1978.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

This work was supported in part by grants #A119811 and #HL25378, Project 1B, from the National Institutes of Health, Bethesda, MD.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Helmke, R.J., Boyd, R.L., German, V.F. et al. From growth factor dependence to growth factor responsiveness: The genesis of an alveolar macrophage cell line. In Vitro Cell Dev Biol 23, 567–574 (1987). https://doi.org/10.1007/BF02620974

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02620974

Key words

Navigation