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479 results on '"Takarada, Takeshi"'

5. Efficient Production of Chondrocyte Particles from Human iPSC-Derived Chondroprogenitors Using a Plate-Based Cell Self-Aggregation Technique.

Author

Hanaki, Shojiro, Yamada, Daisuke, Takao, Tomoka, Iwai, Ryosuke, and Takarada, Takeshi

Subjects
TOLUIDINE blue, SEVERE combined immunodeficiency, ARTICULAR cartilage, IMMUNOHISTOCHEMISTRY, EXTRACELLULAR matrix, CARTILAGE regeneration
Abstract

The limited capacity of articular cartilage for self-repair is a critical challenge in orthopedic medicine. Here, we aimed to develop a simplified method of generating chondrocyte particles from human-induced pluripotent stem cell-derived expandable limb-bud mesenchymal cells (ExpLBM) using a cell self-aggregation technique (CAT). ExpLBM cells were induced to form chondrocyte particles through a stepwise differentiation protocol performed on a CAT plate (prevelex-CAT®), which enables efficient and consistent production of an arbitrary number of uniformly sized particles. Histological and immunohistochemical analyses confirmed that the generated chondrocyte particles expressed key cartilage markers, such as type II collagen and aggrecan, but not hypertrophic markers, such as type X collagen. Additionally, when these particles were transplanted into osteochondral defects in rats with X-linked severe combined immunodeficiency, they demonstrated successful engraftment and extracellular matrix production, as evidenced by Safranin O and Toluidine Blue staining. These data suggest that the plate-based CAT system offers a robust and scalable approach to produce a large number of chondrocyte particles in a simplified and efficient manner, with potential application to cartilage regeneration. Future studies will focus on refining the system and exploring its clinical applications to the treatment of cartilage defects. [ABSTRACT FROM AUTHOR]

Published
2024
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6. Enzyme‐Cleaved Bone Marrow Transplantation Improves the Engraftment of Bone Marrow Mesenchymal Stem Cells

Author

Hotaka Kawai, May Wathone Oo, Kiyofumi Takabatake, Ikue Tosa, Yamin Soe, Htoo Shwe Eain, Sho Sanou, Shigeko Fushimi, Shintaro Sukegawa, Keisuke Nakano, Takarada Takeshi, and Hitoshi Nagatsuka

Subjects
BONE FORMATION, BONE MARROW MESENCHYMAL STEM CELLS, BONE MARROW TRANSPLANTATION MODEL, OSTEOBLASTS, SYSTEM BIOLOGY—BONE INTERACTOR, Orthopedic surgery, RD701-811, Diseases of the musculoskeletal system, RC925-935
Abstract

ABSTRACT Mesenchymal stem cell (MSC) therapy is a promising approach to curing bone diseases and disorders. In treating genetic bone disorders, MSC therapy is local or systemic transplantation of isolated and in vitro proliferated MSC rather than bone marrow transplantation. Recent evidence showed that bone marrow MSC engraftment to bone regeneration has been controversial in animal and human studies. Here, our modified bone marrow transplantation (BMT) method solved this problem. Like routine BMT, our modified method involves three steps: (i) isolation of bone marrow cells from the donor, (ii) whole‐body lethal irradiation to the recipient, and (iii) injection of isolated bone marrow cells into irradiated recipient mice via the tail vein. The significant modification is imported at the bone marrow isolation step. While the bone marrow cells are flushed out from the bone marrow with the medium in routine BMT, we applied the enzymes’ (collagenase type 4 and dispase) integrated medium to wash out the bone marrow cells. Then, cells were incubated in enzyme integrated solution at 37°C for 10 minutes. This modification designated BMT as collagenase‐integrated BMT (c‐BMT). Notably, successful engraftment of bone marrow MSC to the new bone formation, such as osteoblasts and chondrocytes, occurs in c‐BMT mice, whereas routine BMT mice do not recruit bone marrow MSC. Indeed, flow cytometry data showed that c‐BMT includes a higher proportion of LepR+, CD51+, or RUNX2+ non‐hematopoietic cells than BMT. These findings suggested that c‐BMT is a time‐efficient and more reliable technique that ensures the disaggregation and collection of bone marrow stem cells and engraftment of bone marrow MSC to the recipient. Hence, we proposed that c‐BMT might be a promising approach to curing genetic bone disorders. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Published
2023
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9. Antitumor activity of α‐pinene in T‐cell tumors.

Author

Abe, Masaya, Asada, Noboru, Kimura, Maiko, Fukui, Chie, Yamada, Daisuke, Wang, Ziyi, Miyake, Masayuki, Takarada, Takeshi, Ono, Mitsuaki, Aoe, Michinori, Kitamura, Wataru, Matsuda, Masayuki, Moriyama, Takashi, Matsumura, Akifumi, and Maeda, Yoshinobu

Abstract

T‐cell acute leukemia and lymphoma have a poor prognosis. Although new therapeutic agents have been developed, their therapeutic effects are suboptimal. α‐Pinene, a monoterpene compound, has an antitumor effect on solid tumors; however, few comprehensive investigations have been conducted on its impact on hematologic malignancies. This report provides a comprehensive analysis of the potential benefits of using α‐pinene as an antitumor agent for the treatment of T‐cell tumors. We found that α‐pinene inhibited the proliferation of hematologic malignancies, especially in T‐cell tumor cell lines EL‐4 and Molt‐4, induced mitochondrial dysfunction and reactive oxygen species accumulation, and inhibited NF‐κB p65 translocation into the nucleus, leading to robust apoptosis in EL‐4 cells. Collectively, these findings suggest that α‐pinene has potential as a therapeutic agent for T‐cell malignancies, and further investigation is warranted. [ABSTRACT FROM AUTHOR]

Published
2024
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28. Identification of Surface Antigens That Define Human Pluripotent Stem Cell-Derived PRRX1+Limb-Bud-like Mesenchymal Cells

Author

Yamada, Daisuke, Takao, Tomoka, Nakamura, Masahiro, Kitano, Toki, Nakata, Eiji, Takarada, Takeshi, Yamada, Daisuke, Takao, Tomoka, Nakamura, Masahiro, Kitano, Toki, Nakata, Eiji, and Takarada, Takeshi

Abstract

Stem cell-based therapies and experimental methods rely on efficient induction of human pluripotent stem cells (hPSCs). During limb development, the lateral plate mesoderm (LPM) produces limb-bud mesenchymal (LBM) cells that differentiate into osteochondroprogenitor cells and form cartilage tissues in the appendicular skeleton. Previously, we generated PRRX1-tdTomato reporter hPSCs to establish the protocol for inducing the hPSC-derived PRRX1(+) LBM-like cells. However, surface antigens that assess the induction efficiency of hPSC-derived PRRX1(+) LBM-like cells from LPM have not been identified. Here, we used PRRX1-tdTomato reporter hPSCs and found that high pluripotent cell density suppressed the expression of PRRX1 mRNA and tdTomato after LBM-like induction. RNA sequencing and flow cytometry suggested that PRRX1-tdTomato(+) LBM-like cells are defined as CD44(high) CD140B(high) CD49f(-). Importantly, other hPSC lines, including four human induced pluripotent stem cell lines (414C2, 1383D2, HPS1042, HPS1043) and two human embryonic stem cell lines (SEES4, SEES7), showed the same results. Thus, an appropriate cell density of hPSCs before differentiation is a prerequisite for inducing the CD44(high) CD140B(high) CD49f(-) PRRX1(+) LBM-like cells.

Published
2022

29. A protocol to induce expandable limb-bud mesenchymal cells from human pluripotent stem cells

Author

Takao, Tomoka, Yamada, Daisuke, Takarada, Takeshi, Takao, Tomoka, Yamada, Daisuke, and Takarada, Takeshi

Abstract

Here, we present a protocol for the selective differentiation of human pluripotent stem cells mimicking human developmental processes into expandable PRRX1+ limb-bud mesenchymal (ExpLBM) cells. This approach enables expansion through serial passage while maintaining capacity for chondrogenic differentiation. For complete details on the use and execution of this protocol, please refer to Yamada et al. (2021, 2022).

Published
2022

35. Mouse Model for Optogenetic Genome Engineering

Author

Takao, Tomoka, Yamada, Daisuke, and Takarada, Takeshi

Subjects
Optogenetics, Disease Models, Animal, Mice, Animals, Cre recombinase, Genetic Engineering
Abstract

Optogenetics, a technology to manipulate biological phenomena thorough light, has attracted much attention in neuroscience. Recently, the Magnet System, a photo-inducible protein dimerization system which can control the intracellular behavior of various biomolecules with high accuracy using light was developed. Furthermore, photoactivation systems for controlling biological phenomena are being developed by combining this technique with genome-editing technology (CRISPR/Cas9 System) or DNA recombination technology (Cre-loxP system). Herein, we review the history of optogenetics and the latest Magnet System technology and introduce our recently developed photoactivatable Cre knock-in mice with temporal-, spatial-, and cell-specific accuracy.

Published
2022

43. Transcription Cofactor Vestigial-Like 3 Plays an Important Role in Osteoblast Differentiation by Regulating the Expression of Osteogenic Transcription Factors and BMP Signaling

Author

Yuan, Haoze, primary, Ikegame, Mika, additional, Fukuhara, Yoko, additional, Takemoto, Fumiko, additional, Yu, Yaqiong, additional, Weng, Yao, additional, Guo, Jiajie, additional, Yamada, Daisuke, additional, Takarada, Takeshi, additional, Li, Ying, additional, Okamura, Hirohiko, additional, and Zhang, Bin, additional

Published
2022
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