A Stro-1+ human universal stromal feeder layer to expand/maintain human bone marrow hematopoietic stem/progenitor cells in a serum-free culture system

Gonçalves, Raquel; Lobato da Silva, Cláudia; Cabral, Joaquim M.S.; Zanjani, Esmail D.; Almeida-Porada, Graça

doi:10.1016/j.exphem.2006.05.024

« Previous Next »Experimental Hematology
Volume 34, Issue 10 , Pages 1353-1359, October 2006

A Stro-1⁺ human universal stromal feeder layer to expand/maintain human bone marrow hematopoietic stem/progenitor cells in a serum-free culture system

Raquel Gonçalves
- Department of Animal Biotechnology, University of Nevada, Reno, Nev., USA
- Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Lisboa, Portugal
- These authors contributed equally to this work.
Cláudia Lobato da Silva
- Department of Animal Biotechnology, University of Nevada, Reno, Nev., USA
- Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Lisboa, Portugal
- These authors contributed equally to this work.
Joaquim M.S. Cabral
- Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Lisboa, Portugal
Esmail D. Zanjani
- Department of Animal Biotechnology, University of Nevada, Reno, Nev., USA
Graça Almeida-Porada
- Department of Animal Biotechnology, University of Nevada, Reno, Nev., USA
- Offprint requests to: Graça Almeida-Porada, M.D., Ph.D., Department of Animal Biotechnology, University of Nevada, Reno, Mail Stop 202, Reno NV 89557-0104

Received 15 May 2006; accepted 15 May 2006.

Objective

To compare the ability of allogeneic versus autologous purified human Stro-1⁺ mesenchymal stem cell (MSC) populations from different human donors to support the ex vivo expansion and maintenance of human hematopoietic stem/progenitor cells (HSCs). Furthermore, we compared the results obtained with MSC as a feeder layer to traditional allogeneic stromal layers grown in long-term bone marrow culture media (LT-ST).

Methods

Adult human bone marrow CD34⁺-enriched cells were cultured in serum-free medium for 2 to 3 weeks over the respective MSC-irradiated feeder layers or over traditional allogeneic LT- ST stromal layers in the presence of stem cell factor, basic fibroblast growth factor, leukemia inhibitory factor, and Flt-3 and analyzed every 2 to 4 days for expansion, phenotype, and clonogenic ability.

Results

There was a progressive expansion of total numbers of cells in all the experimental groups; however, allogeneic MSCs were more efficient at expanding CD34⁺CD38⁻ cells and showed a higher clonogenic potential than both allogeneic LT-ST and autologous MSCs. The differentiative potential of cells cultured on both MSC and LT-ST was primarily shifted toward myeloid lineage; however, only MSCs were able to maintain/expand a CD7⁺ population with lymphocytic potential. Importantly, transplantation into preimmune fetal sheep demonstrated that the HSCs cultured over MSCs retained their engraftment capability.

Conclusion

These results indicate that purified Stro-1⁺ MSCs may be used as a universal and reproducible stromal feeder layer to efficiently expand and maintain human bone marrow HSCs ex vivo.