| Project/Area Number |
23K17204
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 90110:Biomedical engineering-related
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| Research Institution | Institute of Physical and Chemical Research |
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Principal Investigator |
Koh IsabelSiewYin 国立研究開発法人理化学研究所, 開拓研究本部, 特別研究員 (90868415)
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| Project Period (FY) |
2023-04-01 – 2025-03-31
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| Project Status |
Granted (Fiscal Year 2023)
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| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2024: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
Fiscal Year 2023: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | Neural tube / Ventralisation / Spinal cord / Muti-organ / In vitro modelling |
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| Outline of Research at the Start |
本研究課題の目標は、脳、脊髄、筋肉を感覚神経と運動神経それぞれが独立して結ばれた多臓器神経系モデルの構築を行うこととする。上記目的を達成するため、本研究課題では、①iPS細胞周りの分化因子勾配を制御することにより、感覚および運動神経がそれぞれ局在する脊髄オルガノイドモデルの構築、②脊髄オルガノイドからの感覚および運動神経がそれぞれ伸長し、筋肉オルガノイドへと接続した脊髄－筋肉神経ネットワークの構築、③脳、脊髄、筋肉のオルガノイドが、独立した感覚神経・運動神経により接続された、in vitro多臓器神経系を再構築して、感覚神経および運動神経がそれぞれ解析可能であることを立証することで達成する。
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| Outline of Annual Research Achievements |
An objective of this research is to generate an organoid with motor and sensory neuron components to mimic the spinal cord. To achieve this, an iPSC spheroid was placed in a cube device and cultured in a gradient chip with dorsal and ventral media on opposing ends of the cube to simulate neural tube formation by ventralising signals secreted by the notochord. With this, the neural tube-like structure with localisation of Nkx2.2, a ventral marker, was achieved. The result of this research has shown that culture with localised gradient can lead to localised differentiation, which could provide a method to control the differentiation of cells to the desired pattern, for example in dorsal and ventral localisation, which is important to differentiate to sensory and motor neurons, respectively.
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| Current Status of Research Progress |
Current Status of Research Progress
3: Progress in research has been slightly delayed.
Reason
Although ventral localisation could be observed in the neural tube-like structure by differentiating with gradient culture, dorsal differentiation could not be observed. To overcome this, the concentration of signalling molecules as well as timing and duration of differentiation induction is currently being optimized to obtain neural tube structure with dorsal and ventral regions that will give rise to motor and sensory neurons. At the same time, the method to create two channels in the cube device which will allow neurons from the organoid to elongate out of the cube is currently also being optimised.
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| Strategy for Future Research Activity |
After confirmation of motor-sensory differentiation of the spinal cord organoid, the next objective of this research is to connect the spinal cord organoid to a brain organoid and muscle organoid. Several protocols too generate these organoids have already been reported elsewhere, but the right protocol needs to be confirmed to suit the aims of this research. Furthermore, the media composition and timing and to incorporate three organoids together need to be optimised to suit the co-culture platform being developed in this research project, given the different culture conditions and requirements for each organoid.
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