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Ex vivo expansion of human mobilized peripheral blood stem cells using epigenetic modifiers
Author(s) -
Saraf Santosh,
Araki Hiroto,
Petro Benjamin,
Park Youngmin,
Taioli Simona,
Yoshinaga Kazumi G.,
Koca Emre,
Rondelli Damiano,
Mahmud Nadim
Publication year - 2015
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/trf.12904
Subject(s) - trichostatin a , haematopoiesis , stem cell , ex vivo , biology , microbiology and biotechnology , in vivo , immunology , chemistry , cancer research , histone deacetylase , biochemistry , histone , genetics , gene
Background Epigenetic modifications likely control the fate of hematopoietic stem cells ( HSCs ). The chromatin‐modifying agents ( CMAs ), 5‐aza‐2′‐deoxycytidine (5aza D ) and trichostatin A ( TSA ), have previously been shown to expand HSCs from cord blood and marrow. Here we assessed whether CMA can also expand HSC s present in growth factor–mobilized human peripheral blood ( MPB ). Study Design and Methods 5aza D and TSA were sequentially added to CD 34+ MPB cells in the presence of cytokines, and the cells were cultured for 9 days. Results After culture, a 3.6 ± 0.5‐fold expansion of CD 34+ CD 90+ cells, a 10.1 ± 0.5‐fold expansion of primitive colony‐forming unit ( CFU )‐mix, and a 2.2 ± 0.5‐fold expansion of long‐term cobblestone‐area–forming cells ( CAFCs ) was observed in 5aza D / TSA ‐expanded cells. By contrast, cells cultured in cytokines without 5aza D / TSA displayed no expansion; rather, a reduction in CD 34+ CD 90+ cells (0.7 ± 0.1‐fold) and CAFC s (0.3 ± 0.1‐fold) from their initial numbers was observed. Global hypomethylation corresponding with increased transcript levels of several genes implicated in HSC self‐renewal, including HOXB4 , GATA2 , and EZH2 , was observed in 5aza D / TSA ‐expanded MPB cells in contrast to controls. 5aza D / TSA ‐expanded MPB cells retained in vivo hematopoietic engraftment capacity. Conclusion MPB CD 34+ cells from donors can be expanded using 5aza D / TSA , and these expanded cells retain in vivo hematopoietic reconstitution capacity. This strategy may prove to be potentially useful to augment HSC numbers for patients who fail to mobilize.