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16 results on '"Umekage, Masafumi"'

4. Complete suspension culture of human induced pluripotent stem cells supplemented with suppressors of spontaneous differentiation

Author

Matsuo-Takasaki, Mami, primary, Kambayashi, Sho, additional, Hemmi, Yasuko, additional, Wakabayashi, Tamami, additional, Shimizu, Tomoya, additional, An, Yuri, additional, Takeuchi, Kazuhiro, additional, Ibuki, Masato, additional, Kawai, Yoshikazu, additional, Umekage, Masafumi, additional, Kato, Tomoaki M, additional, Noguchi, Michiya, additional, Nakade, Koji, additional, Nakamura, Yukio, additional, Nakaishi, Tomoyuki, additional, Tsukahara, Masayoshi, additional, and Hayashi, Yohei, additional

Published
2023
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7. Supplement to: Autologous induced stem-cell-derived retinal cells for macular degeneration.

Author

Mandai, Michiko, Watanabe, Akira, Kurimoto, Yasuo, Hirami, Yasuhiko, Morinaga, Chikako, Daimon, Takashi, Fujihara, Masashi, Akimaru, Hiroshi, Sakai, Noriko, Shibata, Yumiko, Terada, Motoki, Nomiya, Yui, Tanishima, Shigeki, Nakamura, Masahiro, Kamao, Hiroyuki, Sugita, Sunao, Onishi, Akishi, Ito, Tomoko, Fujita, Kanako, Kawamata, Shin, Go, Masahiro J., Shinohara, Chikara, Hata, Ken-ichiro, Sawada, Masanori, Yamamoto, Midori, Ohta, Sachiko, Ohara, Yasuo, Yoshida, Kenichi, Kuwahara, Junko, Kitano, Yuko, Amano, Naoki, Umekage, Masafumi, Kitaoka, Fumiyo, Tanaka, Azusa, Okada, Chihiro, Takasu, Naoko, Ogawa, Seishi, Yamanaka, Shinya, and Takahashi, Masayo

Published
2017

8. Generation of hypoimmunogenic induced pluripotent stem cells by CRISPR-Cas9 system and detailed evaluation for clinical application

Author

50578002, Kitano, Yuko, Nishimura, Sayaka, Kato, Tomoaki M., Ueda, Anna, Takigawa, Kaho, Umekage, Masafumi, Nomura, Masaki, Kawakami, Ayane, Ogawa, Haruna, Xu, Huaigeng, Hotta, Akitsu, Takasu, Naoko, Tsukahara, Masayoshi, 50578002, Kitano, Yuko, Nishimura, Sayaka, Kato, Tomoaki M., Ueda, Anna, Takigawa, Kaho, Umekage, Masafumi, Nomura, Masaki, Kawakami, Ayane, Ogawa, Haruna, Xu, Huaigeng, Hotta, Akitsu, Takasu, Naoko, and Tsukahara, Masayoshi

Abstract

In order to expand the promise of regenerative medicine using allogeneic induced pluripotent stem cells (iPSCs), precise and efficient genome editing of human leukocyte antigen (HLA) genes would be advantageous to minimize the immune rejection caused by mismatches of HLA type. However, clinical-grade genome editing of multiple HLA genes in human iPSC lines remains unexplored. Here, we optimized the protocol for good manufacturing practice (GMP)-compatible CRISPR-Cas9 genome editing to deplete the three gene locus (HLA-A, HLA-B, and CIITA genes) simultaneously in HLA homozygous iPSCs. The use of HLA homozygous iPSCs has one main advantage over heterozygous iPSCs for inducing biallelic knockout by a single gRNA. RNA-seq and flow cytometry analyses confirmed the successful depletion of HLAs, and lineage-specific differentiation into cardiomyocytes was verified. We also confirmed that the pluripotency of genome-edited iPSCs was successfully maintained by the three germ layers of differentiation. Moreover, whole-genome sequencing, karyotyping, and optical genome mapping analyses revealed no evident genomic abnormalities detected in some clones, whereas unexpected copy number losses, chromosomal translocations, and complex genomic rearrangements were observed in other clones. Our results indicate the importance of multidimensional analyses to ensure the safety and quality of the genome-edited cells. The manufacturing and assessment pipelines presented here will be the basis for clinical-grade genome editing of iPSCs.

Published
2022

9. Autologous Induced Stem-Cell–Derived Retinal Cells for Macular Degeneration: Brief Report

Author

Mandai, Michiko, Watanabe, Akira, Kurimoto, Yasuo, Hirami, Yasuhiko, Morinaga, Chikako, Daimon, Takashi, Fujihara, Masashi, Akimaru, Hiroshi, Sakai, Noriko, Shibata, Yumiko, Terada, Motoki, Nomiya, Yui, Tanishima, Shigeki, Nakamura, Masahiro, Kamao, Hiroyuki, Sugita, Sunao, Onishi, Akishi, Ito, Tomoko, Fujita, Kanako, Kawamata, Shin, Go, Masahiro J., Shinohara, Chikara, Hata, Ken-ichiro, Sawada, Masanori, Yamamoto, Midori, Ohta, Sachiko, Ohara, Yasuo, Yoshida, Kenichi, Kuwahara, Junko, Kitano, Yuko, Amano, Naoki, Umekage, Masafumi, Kitaoka, Fumiyo, Tanaka, Azusa, Okada, Chihiro, Takasu, Naoko, Ogawa, Seishi, Yamanaka, Shinya, and Takahashi, Masayo

Published
2017
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11. Pre-clinical study of induced pluripotent stem cell-derived dopaminergic progenitor cells for Parkinson’s disease

Author

10587851, 40619821, 10270779, Doi, Daisuke, Magotani, Hiroaki, Kikuchi, Tetsuhiro, Ikeda, Megumi, Hiramatsu, Satoe, Yoshida, Kenji, Amano, Naoki, Nomura, Masaki, Umekage, Masafumi, Morizane, Asuka, Takahashi, Jun, 10587851, 40619821, 10270779, Doi, Daisuke, Magotani, Hiroaki, Kikuchi, Tetsuhiro, Ikeda, Megumi, Hiramatsu, Satoe, Yoshida, Kenji, Amano, Naoki, Nomura, Masaki, Umekage, Masafumi, Morizane, Asuka, and Takahashi, Jun

Abstract

Induced pluripotent stem cell (iPSC)-derived dopaminergic (DA) neurons are an expected source for cell-based therapies for Parkinson’s disease (PD). The regulatory criteria for the clinical application of these therapies, however, have not been established. Here we show the results of our pre-clinical study, in which we evaluate the safety and efficacy of dopaminergic progenitors (DAPs) derived from a clinical-grade human iPSC line. We confirm the characteristics of DAPs by in vitro analyses. We also verify that the DAP population include no residual undifferentiated iPSCs or early neural stem cells and have no genetic aberration in cancer-related genes. Furthermore, in vivo studies using immunodeficient mice reveal no tumorigenicity or toxicity of the cells. When the DAPs are transplanted into the striatum of 6-OHDA-lesioned rats, the animals show behavioral improvement. Based on these results, we started a clinical trial to treat PD patients in 2018.

Published
2020

16. Complete suspension culture of human induced pluripotent stem cells supplemented with suppressors of spontaneous differentiation.

Author

Matsuo-Takasaki M, Kambayashi S, Hemmi Y, Wakabayashi T, Shimizu T, An Y, Ito H, Takeuchi K, Ibuki M, Kawashima T, Masayasu R, Suzuki M, Kawai Y, Umekage M, Kato TM, Noguchi M, Nakade K, Nakamura Y, Nakaishi T, Nishishita N, Tsukahara M, and Hayashi Y

Subjects
Humans, Leukocytes, Mononuclear cytology, Induced Pluripotent Stem Cells cytology, Cell Differentiation, Cell Culture Techniques methods
Abstract

Human induced pluripotent stem cells (hiPSCs) are promising resources for producing various types of tissues in regenerative medicine; however, the improvement in a scalable culture system that can precisely control the cellular status of hiPSCs is needed. Utilizing suspension culture without microcarriers or special materials allows for massive production, automation, cost-effectiveness, and safety assurance in industrialized regenerative medicine. Here, we found that hiPSCs cultured in suspension conditions with continuous agitation without microcarriers or extracellular matrix components were more prone to spontaneous differentiation than those cultured in conventional adherent conditions. Adding PKCβ and Wnt signaling pathway inhibitors in the suspension conditions suppressed the spontaneous differentiation of hiPSCs into ectoderm and mesendoderm, respectively. In these conditions, we successfully completed the culture processes of hiPSCs, including the generation of hiPSCs from peripheral blood mononuclear cells with the expansion of bulk population and single-cell sorted clones, long-term culture with robust self-renewal characteristics, single-cell cloning, direct cryopreservation from suspension culture and their successful recovery, and efficient mass production of a clinical-grade hiPSC line. Our results demonstrate that precise control of the cellular status in suspension culture conditions paves the way for their stable and automated clinical application., Competing Interests: MM, YH Inventor of patents arising from this work (WO/2021/162090 and PCT/JP2020/005255), SK, KT, MI Was employed with KANEKA corporation during this study. Inventor of patents arising from this work (WO/2021/162090 and PCT/JP2020/005255), YH, TW, TS, YA, HI, MU, TK, MN, KN, YN, MT No competing interests declared, TK, RM, MS, YK, TN, NN Was employed with KANEKA corporation during this study, (© 2023, Matsuo-Takasaki et al.)

Published
2024
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