Retroviral vector-mediated gene transfer into primitive human hematopoietic progenitor cells: Effects of mast cell growth factor (MGF) combined with other cytokines

Jan Nolta, G. M. Crooks, R. W. Overell, D. E. Williams, D. B. Kohn

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Retroviral vector-mediated gene transfer into human hematopoietic stem cells may permit gene therapy of numerous genetic diseases. Stimulation of marrow with hematopoietic growth factors (HGFs) has been shown to increase the level of retroviral transduction. We have examined the effects of recombinant human mast cell growth factor (MGF), alone and in combination with other HGFs, on the efficiency of gene transfer into human hematopoietic progenitor cells. MGF acts in concert with interleukin 3 (IL-3) and interleukin 6 (IL-6) to increase the percentage of CD34+ progenitors transduced with a retroviral vector expressing the neo gene. The most potent combination of growth factors that we examined, interleukin 1 (IL-1)/IL-3/IL-6/MGF, resulted in the conferral of G418 resistance to 45% of progenitors and long-term culture-initiating cells. Extending the time of cocultivation of the marrow cells with the vector-producing cells did not further increase gene transfer frequency, suggesting that the amount of available vector is not limiting. To analyze the effects of the HGF on gene transfer into more primitive hematopoietic progenitors, CD34+ cells were isolated from marrow samples that were purged of committed progenitor cells by treatment with 4-hydroperoxycyclophosphamide (4-HC). Preculturing the CD34+ 4-HC-treated cells with the combination of four HGF (IL-1/IL-3/IL-6/MGF) permitted transduction of 20%-28% of the progenitors that formed colonies after 30 days in culture. These results demonstrate that MGF in combination with other HGFs enhances gene transduction of human hematopoietic progenitor cells.

Original languageEnglish (US)
Pages (from-to)1065-1071
Number of pages7
JournalExperimental Hematology
Volume20
Issue number9
StatePublished - 1992
Externally publishedYes

Fingerprint

Stem Cell Factor
Hematopoietic Stem Cells
Intercellular Signaling Peptides and Proteins
perfosfamide
Cytokines
Interleukin-3
Genes
Interleukin-6
Bone Marrow
Interleukin-1
Inborn Genetic Diseases
Coculture Techniques
Gene Frequency
Genetic Therapy
Stem Cells
Cell Culture Techniques

Keywords

  • gene therapy
  • hematopoietic growth factors
  • human hematopoietic progenitor cells
  • mast cell growth factor
  • retroviral vector-mediated gene transfer

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

Retroviral vector-mediated gene transfer into primitive human hematopoietic progenitor cells : Effects of mast cell growth factor (MGF) combined with other cytokines. / Nolta, Jan; Crooks, G. M.; Overell, R. W.; Williams, D. E.; Kohn, D. B.

In: Experimental Hematology, Vol. 20, No. 9, 1992, p. 1065-1071.

Research output: Contribution to journalArticle

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