A novel protocol that allows short-term stem cell expansion of both committed and pluripotent hematopoietic progenitor cells suitable for clinical use
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- 作者：Giuseppe Astori ; Walter Malangone ; Valentina Adami ; Angela Risso ; Laura Dorotea ; Elisabetta Falasca ; Luisa Marini ; Riccardo Spizzo ; Leonardo Bigi ; Pierguido Sala ; Elio Tonutti ; Franco Biffoni ; Cristina Rinaldi ; Giovanni Del Frate ; Marco Pittino ; Alberto Degrassi
- 关键词：umbilical cord blood ; CD34+ cells ; hematopoietic stem cells ; ex vivo expansion
- 刊名：Blood Cells, Molecules, and Diseases
- 出版年：2001
- 出版时间：July 2001
- 年：2001
- 卷：27
- 期：4
- 页码：715-724
- 全文大小：972 K
文摘
To obtain long-term engraftment and hematopoiesis in myeloablated patients, the cell population used for hematopoietic reconstitution should include a sufficient number of early pluripotent hematopoietic stem cells (HSCs), along with committed cells from the various lineages. For this purpose, the small subset of CD34+ cells purified from different sources must be expanded ex vivo. Since cytokines may induce both proliferation and differentiation, expansion would provide a cell population comprising committed as well as uncommitted cells. Optimization of HSC expansion methods could be obtained by a combination of cytokines able to sustain renewal of pluripotent cells yet endowed with poor differentiation potential. We used variations of the combinations of cytokines describedby Brugger et al. [W. Brugger, S. Heimfels, R. J. Berenson, R. Mertelsmann, and L. Kanz (1995) N. Engl. J. Med. 333, 283–287] andPiacibello et al. [W. Piacibello, F. Sanavio, L. Garetto, A. Severino, D. Bergandi, J. Ferrario, F. Fagioli, M. Berger, and M. Aglietta (1997) Blood 89, 2644–2653] to expand UCB CD34+ cells and monitored proliferation rate and phenotype after 14 days of culture. Several hematopoietic lineage-associated surface antigens were evaluated. Our data show that flt3L and thrombopoietin in combination with IL-3, while sustaining a high CD34+ proliferation rate, provide a relatively low enrichment in very early uncommitted CD34+/CD38- cells. Conversely, in the absence of IL-3, they are less effective in inducing proliferation yet significantly increase the number of CD34+/CD38- cells. A combination of the above protocols, applied simultaneously to aliquots of the same sample, would allow expansion of both committed and pluripotent HSC. This strategy may represent a significant improvement for clinical applications.