| 研究実績の概要 |
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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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
3: やや遅れている
理由
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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| 今後の研究の推進方策 |
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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