Access

You are not currently logged in.

Access your personal account or get JSTOR access through your library or other institution:

login

Log in to your personal account or through your institution.

If You Use a Screen Reader

This content is available through Read Online (Free) program, which relies on page scans. Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.

Retroviral Mediated Gene Transfer in Megakaryocytic Cell Lines

William R. Kiffmeyer, Peter J. Stambrook and Michael A. Lieberman
In Vitro Cellular & Developmental Biology. Animal
Vol. 30A, No. 11 (Nov., 1994), pp. 803-809
Stable URL: http://www.jstor.org/stable/4294332
Page Count: 7
  • Read Online (Free)
  • Subscribe ($19.50)
  • Cite this Item
Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
Retroviral Mediated Gene Transfer in Megakaryocytic Cell Lines
Preview not available

Abstract

There have been no reports, to date, of successful introduction of foreign DNA into committed megakaryocyte precursor cells. We have successfully infected two megakaryocytic cell lines, one a committed cell line (CHRF-288-11) and the other a bipotential cell line (K562), with a retroviral vector containing the bacterial lacZ gene and a neomycin resistance marker. Modification of standard protocols was required for successful infection of the committed megakaryocyte cell line. Presence of the lacZ transgene was demonstrated at the molecular level by polymerase chain reaction (PCR), and its expression at the mRNA level by reverse transcriptase-PCR. Presence of the bacterial β-galactosidase was demonstrated by both immunofluorescence and enzyme activity. Treatment of the CHRF-288-11 infected cells with phorbol esters, which induces megakaryocytic differentiation, increases expression of the lacZ transgene. The staining pattern of the lacZ reaction product was perinuclear and punctate in the CHRF-288-11 cells, whereas it was uniform throughout the cytoplasm of the K562 cells, suggesting different sorting mechanisms for bacterial β-galactosidase in these two cell types. Overall, these results demonstrate the feasibility and provide a method for infecting cultured megakaryocytic cell lines with retroviral of vectors such that a molecular analysis of megakaryocyte differentiation can be accomplished.

Page Thumbnails

  • Thumbnail: Page 
803
    803
  • Thumbnail: Page 
804
    804
  • Thumbnail: Page 
805
    805
  • Thumbnail: Page 
806
    806
  • Thumbnail: Page 
807
    807
  • Thumbnail: Page 
808
    808
  • Thumbnail: Page 
809
    809