{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,11]],"date-time":"2026-03-11T23:28:33Z","timestamp":1773271713501,"version":"3.50.1"},"reference-count":55,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2004,11,10]],"date-time":"2004-11-10T00:00:00Z","timestamp":1100044800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Journal of Gene Medicine"],"published-print":{"date-parts":[[2004,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>One of the major barriers to the clinical application of hematopoietic stem cell (HSC) gene therapy has been relatively low gene transfer efficiency. Other inadequacies of current transduction protocols are related to their multi\u2010step procedures, e.g., using tissue\u2010culture flasks, roller bottles or gas\u2010permeable bags for clinical application.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>In comparison with a conventional bag transduction protocol, a \u2018closed\u2019 hollow\u2010fiber bioreactor system (HBS) was exploited to culture and transduce human peripheral blood CD34<jats:sup>+<\/jats:sup> progenitor cells (PBPC<jats:sub>MPS<\/jats:sub>) from patients with mucopolysaccharidosis type I (MPS I) using an amphotropic retroviral vector based on a murine Moloney leukemia virus LN prototype. Both short\u2010term colony\u2010forming cell (CFC) and long\u2010term culture initiating cell (LTCIC) assays were employed to determine transduction frequency and transgene expression in committed progenitor cells and primitive progenitors with multi\u2010lineage potentials.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>A novel ultrafiltration\u2010transduction method was established to culture and transduce enzyme\u2010deficient PBPC<jats:sub>MPS<\/jats:sub> over a 5\u2010day period without loss in viability and CD34 identity (<jats:italic>n<\/jats:italic> = 5). Significantly higher transduction efficiencies were achieved in primary CFC that derived from the HBS (5.8\u201314.2%) in comparison with those from gas\u2010permeable bags (undetectable to 1.7%; <jats:italic>p<\/jats:italic> &lt; 0.01). Up to 15\u2010fold higher\u2010than\u2010normal enzyme activity was found in selected PBPC<jats:sub>MPS<\/jats:sub>\u2010LP1CD transductants. Moreover, higher gene transfer (4.4\u2010fold) and expression in very primitive progenitors were observed in products from the HBS compared with bag experiments as indicated by CFC derived from primitive LTCIC. Remarkably, with relatively modest gene transfer levels in LTCIC from HBS experiments, the expression of the IDUA transgene corrected the enzyme\u2010deficiency in 5\u2010week long\u2010term cultures (LTC).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>MPS I progenitor cells achieved normalized enzyme levels in LTC after transduction in a HBS system. These studies demonstrate the advantages of a bioreactor\u2010transduction system for viral\u2010mediated stem cell gene transfer. 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