Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice
Increasing demand for human hematopoietic stem cells (HSCs) in clinical and research applications necessitates expansion of HSCs in vitro. Before these cells can be used they must be carefully evaluated to assess their stem cell activity. Here, we expanded cord blood CD34+ CD133+ cells in a defined...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Public Library of Science
2011
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| Online Access: | http://hdl.handle.net/1721.1/66282 https://orcid.org/0000-0002-7029-7415 https://orcid.org/0000-0002-5687-6154 |
| _version_ | 1811073660584198144 |
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| author | Drake, Adam Khoury, Maroun Leskov, Ilya B. Iliopoulou, Bettina P. Fragoso, Maria F. Lodish, Harvey F. Chen, Jianzhu |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Drake, Adam Khoury, Maroun Leskov, Ilya B. Iliopoulou, Bettina P. Fragoso, Maria F. Lodish, Harvey F. Chen, Jianzhu |
| author_sort | Drake, Adam |
| collection | MIT |
| description | Increasing demand for human hematopoietic stem cells (HSCs) in clinical and research applications necessitates expansion of HSCs in vitro. Before these cells can be used they must be carefully evaluated to assess their stem cell activity. Here, we expanded cord blood CD34+ CD133+ cells in a defined medium containing angiopoietin like 5 and insulin-like growth factor binding protein 2 and evaluated the cells for stem cell activity in NOD-SCID Il2rg−/− (NSG) mice by multi-lineage engraftment, long term reconstitution, limiting dilution and serial reconstitution. The phenotype of expanded cells was characterized by flow cytometry during the course of expansion and following engraftment in mice. We show that the SCID repopulating activity resides in the CD34+ CD133+ fraction of expanded cells and that CD34+ CD133+ cell number correlates with SCID repopulating activity before and after culture. The expanded cells mediate long-term hematopoiesis and serial reconstitution in NSG mice. Furthermore, they efficiently reconstitute not only neonate but also adult NSG recipients, generating human blood cell populations similar to those reported in mice reconstituted with uncultured human HSCs. These findings suggest an expansion of long term HSCs in our culture and show that expression of CD34 and CD133 serves as a marker for HSC activity in human cord blood cell cultures. The ability to expand human HSCs in vitro should facilitate clinical use of HSCs and large-scale construction of humanized mice from the same donor for research applications. |
| first_indexed | 2024-09-23T09:36:33Z |
| format | Article |
| id | mit-1721.1/66282 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:36:33Z |
| publishDate | 2011 |
| publisher | Public Library of Science |
| record_format | dspace |
| spelling | mit-1721.1/662822022-09-30T15:38:12Z Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice Drake, Adam Khoury, Maroun Leskov, Ilya B. Iliopoulou, Bettina P. Fragoso, Maria F. Lodish, Harvey F. Chen, Jianzhu Massachusetts Institute of Technology. Department of Biology Singapore-MIT Alliance in Research and Technology (SMART) Whitehead Institute for Biomedical Research Koch Institute for Integrative Cancer Research at MIT Chen, Jianzhu Chen, Jianzhu Drake, Adam Khoury, Maroun Leskov, Ilya B. Iliopoulou, Bettina P. Fragoso, Maria F. Lodish, Harvey F. Increasing demand for human hematopoietic stem cells (HSCs) in clinical and research applications necessitates expansion of HSCs in vitro. Before these cells can be used they must be carefully evaluated to assess their stem cell activity. Here, we expanded cord blood CD34+ CD133+ cells in a defined medium containing angiopoietin like 5 and insulin-like growth factor binding protein 2 and evaluated the cells for stem cell activity in NOD-SCID Il2rg−/− (NSG) mice by multi-lineage engraftment, long term reconstitution, limiting dilution and serial reconstitution. The phenotype of expanded cells was characterized by flow cytometry during the course of expansion and following engraftment in mice. We show that the SCID repopulating activity resides in the CD34+ CD133+ fraction of expanded cells and that CD34+ CD133+ cell number correlates with SCID repopulating activity before and after culture. The expanded cells mediate long-term hematopoiesis and serial reconstitution in NSG mice. Furthermore, they efficiently reconstitute not only neonate but also adult NSG recipients, generating human blood cell populations similar to those reported in mice reconstituted with uncultured human HSCs. These findings suggest an expansion of long term HSCs in our culture and show that expression of CD34 and CD133 serves as a marker for HSC activity in human cord blood cell cultures. The ability to expand human HSCs in vitro should facilitate clinical use of HSCs and large-scale construction of humanized mice from the same donor for research applications. Singapore-MIT Alliance for Research and Technology ( Infectious Diseases research grant) 2011-10-17T16:05:05Z 2011-10-17T16:05:05Z 2011-04 2010-10 Article http://purl.org/eprint/type/JournalArticle 1932-6203 http://hdl.handle.net/1721.1/66282 Drake, Adam C. et al. (2011) "Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice." PLoS ONE 6(4): e18382. https://orcid.org/0000-0002-7029-7415 https://orcid.org/0000-0002-5687-6154 en_US http://dx.doi.org/10.1371/journal.pone.0018382 PLoS One Creative Commons Attribution http://creativecommons.org/licenses/by/2.5/ application/pdf Public Library of Science PLoS |
| spellingShingle | Drake, Adam Khoury, Maroun Leskov, Ilya B. Iliopoulou, Bettina P. Fragoso, Maria F. Lodish, Harvey F. Chen, Jianzhu Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice |
| title | Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice |
| title_full | Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice |
| title_fullStr | Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice |
| title_full_unstemmed | Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice |
| title_short | Human CD34+ CD133+ Hematopoietic Stem Cells Cultured with Growth Factors Including Angptl5 Efficiently Engraft Adult NOD-SCID Il2rγ−/− (NSG) Mice |
| title_sort | human cd34 cd133 hematopoietic stem cells cultured with growth factors including angptl5 efficiently engraft adult nod scid il2rγ nsg mice |
| url | http://hdl.handle.net/1721.1/66282 https://orcid.org/0000-0002-7029-7415 https://orcid.org/0000-0002-5687-6154 |
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