Establishment of induced pluripotent stem cells from normal B cells and inducing AID expression in their differentiation into hematopoietic progenitor cells
| Autor: | Atsushi Katafuchi, Makoto Inaki, Fumihiko Kawamura, Akira Sakai, Shinichi Suzuki, Yu Abe, Takumi Yamaura, Megumi Sasatani, Kenji Kamiya, Yumiko Kurosu, Hiroyuki Suzuki, Satoshi Muto, Hideyoshi Noji, Naohiro Tsuyama, Mitsunori Higuchi, Mitsuaki A. Yoshida, Aki Yanagi, Masafumi Onodera |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Science Antigens CD19 Induced Pluripotent Stem Cells CD34 Mice SCID Biology Models Biological Article Colony-Forming Units Assay 03 medical and health sciences Kruppel-Like Factor 4 SOX2 Cytidine Deaminase medicine Animals Humans Induced pluripotent stem cell B cell B-Lymphocytes Multidisciplinary CD40 Cell Differentiation Gene rearrangement Transfection Hematopoietic Stem Cells Molecular biology Hematopoiesis Endothelial stem cell 030104 developmental biology medicine.anatomical_structure Doxycycline Enzyme Induction embryonic structures biology.protein Medicine Lymph Nodes |
| Zdroj: | Scientific Reports Scientific Reports, Vol 7, Iss 1, Pp 1-11 (2017) |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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