Functional assessment of the engraftment potential of gammaretrovirus- modified CD34 + cells, using a short serum-free transduction protocol Journal Article


Authors: Budak-Alpdogan, T.; Przybylowski, M.; Gonen, M.; Sadelain, M.; Bertino, J.; Riviere, I.
Article Title: Functional assessment of the engraftment potential of gammaretrovirus- modified CD34 + cells, using a short serum-free transduction protocol
Abstract: The successful transduction and engraftment of human mobilized peripheral blood (MBP) CD34 + cells are determined to a large extent by the ex vivo cell-processing conditions. In preparation for upcoming clinical trials, we investigated essential culture parameters and devised a short and efficient gammaretroviral transduction protocol entailing minimal manipulation of MBP CD34 + cells. The engraftment potential and in vivo transgene expression in the progeny of repopulating CD34 + cells were measured to assess the functionality of CD34 + cells transduced under these conditions. Using a competitive in vivo repopulation assay in nonobese diabetic/severe combined immunodeficient mice, we demonstrate equivalent engraftment of CD34 + cells transduced under serum-free conditions as compared with CD34 + cells cultured with serum. We also took advantage of this in vivo model to demonstrate that ex vivo manipulation of CD34 + cells can be shortened to 60 hr, using 36 hr of prestimulation and two cycles of transduction 12 hr apart. These minimally manipulated CD34 + cells engraft in a manner similar to cells transduced under longer protocols and the vector-encoded transgene is expressed at the same frequency in cells derived from repopulating CD34 + cells in vivo. We have thus developed a short and efficient human MBP CD34 + transduction protocol under serum-free conditions that is suitable and broadly applicable for phase I clinical trials. © Mary Ann Liebert, Inc.
Keywords: controlled study; human cell; nonhuman; mouse; animals; mice; cell survival; cell function; cd34 antigen; stem cell factor; gene expression; culture medium; animal experiment; assay; time; genetic transduction; genetic vectors; transduction, genetic; mice, inbred strains; cell culture; gene therapy; progeny; transgene; cell culture techniques; blood cell; blood cells; cattle; tetrahydrofolate dehydrogenase; antigens, cd34; serum; scid mouse; transgenes; cell transplantation; retrovirus; virus vector; gammaretrovirus; competitive repopulation assay; gammaretrovirus vector
Journal Title: Human Gene Therapy
Volume: 17
Issue: 7
ISSN: 1043-0342
Publisher: Mary Ann Liebert, Inc  
Date Published: 2006-07-01
Start Page: 780
End Page: 794
Language: English
DOI: 10.1089/hum.2006.17.780
PUBMED: 16839276
PROVIDER: scopus
DOI/URL:
Notes: --- - "Cited By (since 1996): 5" - "Export Date: 4 June 2012" - "CODEN: HGTHE" - "Source: Scopus"
Altmetric Score
MSK Authors
  1. Mithat Gonen
    713 Gonen
  2. Michel W J Sadelain
    456 Sadelain
  3. Isabelle C Riviere
    164 Riviere