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000034345 1001_ $$0P:(DE-Juel1)VDB694$$aJelinek, Nanni$$b0$$uFZJ
000034345 245__ $$aKultivierung hämatopoetischer Zellen : Kokultur im Festbettreaktor
000034345 260__ $$aJülich$$bForschungszentrum Jülich GmbH Zentralbibliothek, Verlag$$c2000
000034345 300__ $$aX, 192 p.
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000034345 4900_ $$0PERI:(DE-600)2414853-2$$8543$$aBerichte des Forschungszentrums Jülich$$v3863$$x0944-2952
000034345 502__ $$aAachen, Techn. Hochsch., Diss.$$bDr. (FH)$$cTechn. Hochsch. Aachen$$d2000
000034345 500__ $$aRecord converted from VDB: 12.11.2012
000034345 520__ $$aThe ex vivo expansion of hematopoietic cells is an important technique for many medical applications. The cultivation concept in this study was the ex vivo simulation of the human bone marrow by co-culture of hematopoietic and stromal cells in a fixed-bed bioreactor . Thereby the cells are immobilized on macroporous collagen microcarriers. A miniaturized loop-reactor was established for screening purposes. For the full supply with primary CD34<sup>+</sup>- cells a cord blood bank was built up. In preliminary investigations two stromal cellines were characterized and the harvest of immobilized cells established by enzymatic lysis of the carriers. The relevant parameters for the co-culture were optimized in the miniaturized loop- reactor. In this system the mononuclear cells (MNC) were expanded up to 454 fold, the colony-forming-cells (CFC) up to 26 fold and the cobblestone-area-forming cells (CAFC) up to 18 fold. In this system the SCm-mouse repopulating cells (SRC) were maintained in contrast to a parallel cultivation in a suspension system. This demonstrates the positive effect of the stromal based, immobilized culture compared to the suspension culture of this cells. Based on the optimization in the screening system the hematopoietic cells were successfully expanded in the fixed-bed bioreactor. The co-culture with the stromal cellline SVSl and the cytokines IL-3, TPO, SCF and FL led to the best results. In this system the MNC were expanded maximal 103 fold, the CFU-GM 114 fold and the CAFC 15 fold. These data correspond the medical yardstick demonstrating the clinical relevance of the results presented in this study.
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