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Immunocytochemical identification of murine and human megakaryocyte colonies in soft-agar cultures

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Murine megakaryocyte (MK) colonies in soft-agar cultures were immunocytochemically stained with platelet antiserum and an immuno-alkaline phosphatase procedure. Subsequently, cytochemical staining for acetylcholinesterase was used to confirm the specificity of the immunolabelling technique. The correlation of numbers of megakaryocyte colonies enumerated by independent observers was excellent. A comparable platelet antiserum directed against human platelet epitopes was utilized to identify human MK colonies in soft-agar cultures of human bone marrow. Using this method, we determined that the frequency of detectable human MK colonies in our agar culture system was maximal between days 10 and 12. The immunocytochemical staining technique we have developed for identification of MK colonies in soft-agar cultures yielded good cellular morphology and produced an intensely specific label against a clear background; it therefore facilitated accurate enumeration of MK colonies. This non-fluorescent method avoids dependence upon a non-permanent marker, and allows the simultaneous enumeration of positive and negative colonies.

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Authors and Affiliations

  1. Department of Laboratory Medicine, University of California School of Medicine, San Francisco, CA, USA

    Jun Liang Zhang, Georgiann Baker & Jack Levin

  2. The Veterans Administration Medical Center, San Francisco, CA, USA

    Jun Liang Zhang, Georgiann Baker & Jack Levin

  3. Department of Pathology, Oregon Health Sciences University, Portland, OR, USA

    Paula E. Stenberg

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  1. Jun Liang Zhang
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  2. Paula E. Stenberg
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  3. Georgiann Baker
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  4. Jack Levin
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Zhang, J.L., Stenberg, P.E., Baker, G. et al. Immunocytochemical identification of murine and human megakaryocyte colonies in soft-agar cultures. Histochem J 26, 170–178 (1994). https://doi.org/10.1007/BF00157966

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  • DOI: https://doi.org/10.1007/BF00157966

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