Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells

Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells

Abstract B cell derived induced pluripotent stem cells (BiPSCs) were recently established from peripheral blood B cells by the simultaneous transfection of Yamanaka factors (Oct3/4, Sox2, Klf4, c-Myc) and C/EBPα using a Sendai virus vector. Here, using a different method, we established BiPSCs with...

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Autores principales: Fumihiko Kawamura, Makoto Inaki, Atsushi Katafuchi, Yu Abe, Naohiro Tsuyama, Yumiko Kurosu, Aki Yanagi, Mitsunori Higuchi, Satoshi Muto, Takumi Yamaura, Hiroyuki Suzuki, Hideyoshi Noji, Shinichi Suzuki, Mitsuaki A. Yoshida, Megumi Sasatani, Kenji Kamiya, Masafumi Onodera, Akira Sakai
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Publicado: Nature Portfolio 2017
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Acceso en línea:https://doaj.org/article/356c7f8b817b42dab5c51de1994d8612
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spelling oai:doaj.org-article:356c7f8b817b42dab5c51de1994d86122021-12-02T16:06:18ZEstablishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells10.1038/s41598-017-01627-12045-2322https://doaj.org/article/356c7f8b817b42dab5c51de1994d86122017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01627-1https://doaj.org/toc/2045-2322Abstract B cell derived induced pluripotent stem cells (BiPSCs) were recently established from peripheral blood B cells by the simultaneous transfection of Yamanaka factors (Oct3/4, Sox2, Klf4, c-Myc) and C/EBPα using a Sendai virus vector. Here, using a different method, we established BiPSCs with immunoglobulin heavy chain (IgH) gene rearrangement from normal B cells purified from lymph nodes. The critical points of our method are pre-stimulation of B cells with IL-21 and CD40-ligand (CD40L), followed by consecutive transfection of highly concentrated Yamanaka factors using a retroviral vector. Following each transfection the cells were centrifuged onto a retronectin coated plate and the activated by IL-4, IL-2, and CD40L. Furthermore, we established BiPSCs (BiPSC-A) in which activation-induced cytidine deaminase (AID) could be induced using the doxycycline-controlled. Both the parental BiPSC and BiPSC-A showed the capability of differentiating into hematopoietic progenitor cells (HPCs) based on confirmation of CD34 expression and colony-formation from CD34-positive cells. The findings that BiPSC-A can differentiate into HPCs suggest that there is a possibility that induction of AID expression would result in chromosomal translocations in the process of differentiation from BiPSCs, and therefore that these BiPSCs could be useful in elucidating the tumor origin of abnormal B cells in myelomagenesis.Fumihiko KawamuraMakoto InakiAtsushi KatafuchiYu AbeNaohiro TsuyamaYumiko KurosuAki YanagiMitsunori HiguchiSatoshi MutoTakumi YamauraHiroyuki SuzukiHideyoshi NojiShinichi SuzukiMitsuaki A. YoshidaMegumi SasataniKenji KamiyaMasafumi OnoderaAkira SakaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution DOAJ
collection DOAJ
language EN
topic Medicine
R
Science
Q
spellingShingle Medicine
R
Science
Q
Fumihiko Kawamura
Makoto Inaki
Atsushi Katafuchi
Yu Abe
Naohiro Tsuyama
Yumiko Kurosu
Aki Yanagi
Mitsunori Higuchi
Satoshi Muto
Takumi Yamaura
Hiroyuki Suzuki
Hideyoshi Noji
Shinichi Suzuki
Mitsuaki A. Yoshida
Megumi Sasatani
Kenji Kamiya
Masafumi Onodera
Akira Sakai
Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells
description Abstract B cell derived induced pluripotent stem cells (BiPSCs) were recently established from peripheral blood B cells by the simultaneous transfection of Yamanaka factors (Oct3/4, Sox2, Klf4, c-Myc) and C/EBPα using a Sendai virus vector. Here, using a different method, we established BiPSCs with immunoglobulin heavy chain (IgH) gene rearrangement from normal B cells purified from lymph nodes. The critical points of our method are pre-stimulation of B cells with IL-21 and CD40-ligand (CD40L), followed by consecutive transfection of highly concentrated Yamanaka factors using a retroviral vector. Following each transfection the cells were centrifuged onto a retronectin coated plate and the activated by IL-4, IL-2, and CD40L. Furthermore, we established BiPSCs (BiPSC-A) in which activation-induced cytidine deaminase (AID) could be induced using the doxycycline-controlled. Both the parental BiPSC and BiPSC-A showed the capability of differentiating into hematopoietic progenitor cells (HPCs) based on confirmation of CD34 expression and colony-formation from CD34-positive cells. The findings that BiPSC-A can differentiate into HPCs suggest that there is a possibility that induction of AID expression would result in chromosomal translocations in the process of differentiation from BiPSCs, and therefore that these BiPSCs could be useful in elucidating the tumor origin of abnormal B cells in myelomagenesis.
format article
author Fumihiko Kawamura
Makoto Inaki
Atsushi Katafuchi
Yu Abe
Naohiro Tsuyama
Yumiko Kurosu
Aki Yanagi
Mitsunori Higuchi
Satoshi Muto
Takumi Yamaura
Hiroyuki Suzuki
Hideyoshi Noji
Shinichi Suzuki
Mitsuaki A. Yoshida
Megumi Sasatani
Kenji Kamiya
Masafumi Onodera
Akira Sakai
author_facet Fumihiko Kawamura
Makoto Inaki
Atsushi Katafuchi
Yu Abe
Naohiro Tsuyama
Yumiko Kurosu
Aki Yanagi
Mitsunori Higuchi
Satoshi Muto
Takumi Yamaura
Hiroyuki Suzuki
Hideyoshi Noji
Shinichi Suzuki
Mitsuaki A. Yoshida
Megumi Sasatani
Kenji Kamiya
Masafumi Onodera
Akira Sakai
author_sort Fumihiko Kawamura
title Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells
title_short Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells
title_full Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells
title_fullStr Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells
title_full_unstemmed Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells
title_sort establishment of induced pluripotent stem cells from normal b cells and inducing aid expression in their differentiation into hematopoietic progenitor cells
publisher Nature Portfolio
publishDate 2017
url https://doaj.org/article/356c7f8b817b42dab5c51de1994d8612
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