Genetische Markierung von Stammzellen zur Analyse der Herkunft und Klonalitaet von Rezidivzellen nach Knochenmarktransplantation Schlussbericht

We have developed the experimental and logistical conditions for the clinical application of retroviral gene transfer. Culture conditions and duration, growth factors and vector pseudotype have been defined for successful retroviral transduction of primitive human hematopoietic progenitor and stem cells and have been measured in phenotypically and functionally defined populations. A new medium for ex vivo transduction culture of hematopoietic stem cells and a serum free vector production method have been developed and vector production was performed. An internationally competitive clinical CML study on purging and retroviral gene transfer into hematopoietic stem cells under GMP conditions has been performed for the first time in Germany. In addition to molecular assays of biological safety and quality control, an internally controlled assay for exact vector quantification has been developed. Further the group has invented a method for sensitive analysis of clonal vector integration. With this method it is possible to track clones of unique stem cells for the first time in vivo over the course of time. The conditions for carrying out cross-section and longitudinal analyses have been acquired. In case that early detectable transplanted stem cells prove to be essential for longtime hematopoiesis, the achieved efficiency of gene transfer reaches a therapeutic level. (orig.)SIGLEAvailable from TIB Hannover: DtF QN1(81,5) / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Bildung und Forschung (BMBF), Bonn (Germany)DEGerman