| 研究課題/領域番号 |
21K16203
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| 研究機関 | 筑波大学 |
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研究代表者 |
周 思奇 筑波大学, 生命環境系, 研究員 (60869676)
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| 研究期間 (年度) |
2021-04-01 – 2023-03-31
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| キーワード | iPS cell / 3D organoid / hair follicles |
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| 研究実績の概要 |
In this study, the primary objective was to explore whether it is possible to efficiently and accurately model hair follicle development using human induced pluripotent stem cell (iPS cells), and to establish a model of hair loss simulated by applying appropriate exogenous stress.
In the experimental proposal, I mentioned what I will do in the first year, mainly focusing on: 1, the establishment of the iPS cell culture system; 2, the differentiation to dermal papilla cells and hair follicle keratinocytes; 3, Inducing iPS cells to 3D cell organoids, then inducing differentiation process to hair follicle tissue by an in vitro model.
Firstly, I established the culture system after we got iPS cells from Riken cell bank in the last year. By searching papers, a relatively simple culture method has been selected. I have repeated over 10 times subcultures to confirm that this culture system is appropriate for iPS cells culture. And the cells have been verified that still retain the ability of pluripotent differentiation by testing the marker genes of iPS cells after multiple passages in this culture system.
Next, I tried to induce the iPS cells differentiation by the conditions mentioned in the previous study. Initially, I determined the growth factors necessary for differentiation induction, but I have not yet performed the verification of characteristic genes. Also, the iPS cell organoid construction have been started.
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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
Due to the impact of the COVID-19 in the first half of last year, many experimental consumables and culture media were out of stock in Japan or could not be delivered in time. Therefore, there is some delay in the start of the experiment. Fortunately, from the second half of the year, this situation gradually recovered, and the experiment was gradually carried out.
When I started culturing iPS cells, due to some differences between iPS cell and general cell culture conditions, I wasted some time in the process of establishing the culture system. Fortunately, after the establishment of the culture system and the familiarity with the experimental operation, the subsequent experiments can be carried out smoothly.
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| 今後の研究の推進方策 |
The following experiments will mainly focus on the following points.
1. At this stage, we have induced the differentiation of iPS cells into keratinocytes and dermal papilla cells, but the evaluation of cell differentiation and the detection of marker genes for the mature differentiated cells have not yet been determined, so we will prioritize the optimization of cell differentiation conditions.
2. Next, I will conduct the next experiment, the construction of 3D organoid. At present, the construction of this structure is still in the early stage of the experiment. It is still unclear whether the 3D structure can be formed stably, how long it can survive, and whether it can be differentiated. Therefore, we will do further research on the stability of the 3D organoid structure.
3. Finally, within this year, it is expected to start conducting differentiation experiments on 3D organoids, inducing them to form hair follicle structures in vitro., and verifying whether the structures can generate new hairs.
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| 次年度使用額が生じた理由 |
Due to the delay in the start of the experiment, as well as the inventory and distribution problems of various commodities caused by COVID-19, I used more consumables that were reserved before. Therefore, there is an amount of money that cannot be used up.
Starting this year, I will use this money for more culture medium , antibodies and assay kits for the further experiments. In the next experiment, I first need to optimize the conditions of cell differentiation, so I need to purchase some corresponding growth factors. At the same time, in order to verify the markers of differentiated cells, I also need to purchase the corresponding antibodies for verification. Second, in order to build 3D structures, I need specially well-plates for culturing. Previously, I was still using the plates that we had stocked up in the past. Starting this year, I will need to repurchase them again. Finally, in the 3D hair follicle induction and differentiation experiment, I need to purchase some special kits or growth factors. In addition, in order to verify the structure of the hair follicle, staining assay is required. Therefore, some special antibodies need to be purchased.
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