Decitabine suspends human CD34+ cell differentiation and proliferation during lentiviral transduction.
Efficient ex vivo transduction of hematopoietic stem cells (HSCs) is encumbered by differentiation which reduces engraftment. We hypothesized that inhibiting DNA methyltransferase with decitabine would block differentiation of transduced CD34+ cells under cytokine stimulation and thus improve transd...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4121241?pdf=render |
| _version_ | 1818214485441118208 |
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| author | Naoya Uchida Matthew M Hsieh Charlotte Platner Yogen Saunthararajah John F Tisdale |
| author_facet | Naoya Uchida Matthew M Hsieh Charlotte Platner Yogen Saunthararajah John F Tisdale |
| author_sort | Naoya Uchida |
| collection | DOAJ |
| description | Efficient ex vivo transduction of hematopoietic stem cells (HSCs) is encumbered by differentiation which reduces engraftment. We hypothesized that inhibiting DNA methyltransferase with decitabine would block differentiation of transduced CD34+ cells under cytokine stimulation and thus improve transduction efficiency for engrafting HSCs. Human CD34+ cells in cytokine-containing media were treated with or without decitabine for 24 or 48 hours, and then these cells were transduced with a GFP-expressing lentiviral vector. Utilizing decitabine pre-treatment for 48 hours, we observed an equivalent percentage of successfully transduced cells (GFP-positivity) and a higher percentage of cells that retained CD34 positivity, compared to no decitabine exposure. Cell proliferation was inhibited after decitabine exposure. Similar results were observed among CD34+ cells from six different donors. Repopulating activity was evaluated by transplantation into NOD/SCID/IL2Rγnull mice and demonstrated an equivalent percentage of GFP-positivity in human cells from decitabine-treated samples and a trend for higher human cell engraftment (measured 20-24 weeks after transplantation), compared to no decitabine exposure. In conclusion, ex vivo decitabine exposure inhibits both differentiation and proliferation in transduced human CD34+ cells and modestly increases the engraftment ability in xenograft mice, while the transduction efficiency is equivalent in decitabine exposure, suggesting improvement of lentiviral transduction for HSCs. |
| first_indexed | 2024-12-12T06:20:56Z |
| format | Article |
| id | doaj.art-72e12464501a4973ace3fece1286ea51 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T06:20:56Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-72e12464501a4973ace3fece1286ea512022-12-22T00:34:55ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0198e10402210.1371/journal.pone.0104022Decitabine suspends human CD34+ cell differentiation and proliferation during lentiviral transduction.Naoya UchidaMatthew M HsiehCharlotte PlatnerYogen SaunthararajahJohn F TisdaleEfficient ex vivo transduction of hematopoietic stem cells (HSCs) is encumbered by differentiation which reduces engraftment. We hypothesized that inhibiting DNA methyltransferase with decitabine would block differentiation of transduced CD34+ cells under cytokine stimulation and thus improve transduction efficiency for engrafting HSCs. Human CD34+ cells in cytokine-containing media were treated with or without decitabine for 24 or 48 hours, and then these cells were transduced with a GFP-expressing lentiviral vector. Utilizing decitabine pre-treatment for 48 hours, we observed an equivalent percentage of successfully transduced cells (GFP-positivity) and a higher percentage of cells that retained CD34 positivity, compared to no decitabine exposure. Cell proliferation was inhibited after decitabine exposure. Similar results were observed among CD34+ cells from six different donors. Repopulating activity was evaluated by transplantation into NOD/SCID/IL2Rγnull mice and demonstrated an equivalent percentage of GFP-positivity in human cells from decitabine-treated samples and a trend for higher human cell engraftment (measured 20-24 weeks after transplantation), compared to no decitabine exposure. In conclusion, ex vivo decitabine exposure inhibits both differentiation and proliferation in transduced human CD34+ cells and modestly increases the engraftment ability in xenograft mice, while the transduction efficiency is equivalent in decitabine exposure, suggesting improvement of lentiviral transduction for HSCs.http://europepmc.org/articles/PMC4121241?pdf=render |
| spellingShingle | Naoya Uchida Matthew M Hsieh Charlotte Platner Yogen Saunthararajah John F Tisdale Decitabine suspends human CD34+ cell differentiation and proliferation during lentiviral transduction. PLoS ONE |
| title | Decitabine suspends human CD34+ cell differentiation and proliferation during lentiviral transduction. |
| title_full | Decitabine suspends human CD34+ cell differentiation and proliferation during lentiviral transduction. |
| title_fullStr | Decitabine suspends human CD34+ cell differentiation and proliferation during lentiviral transduction. |
| title_full_unstemmed | Decitabine suspends human CD34+ cell differentiation and proliferation during lentiviral transduction. |
| title_short | Decitabine suspends human CD34+ cell differentiation and proliferation during lentiviral transduction. |
| title_sort | decitabine suspends human cd34 cell differentiation and proliferation during lentiviral transduction |
| url | http://europepmc.org/articles/PMC4121241?pdf=render |
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