2022 Fiscal Year Final Research Report
Live cell recovery and wide area analysis of dynamic lymphoid tissue subcompartments
| Project/Area Number |
19K22528
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 49:Pathology, infection/immunology, and related fields
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| Research Institution | Niigata University |
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Principal Investigator |
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| Project Period (FY) |
2019-06-28 – 2023-03-31
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| Keywords | 細胞単離 / ストローマ細胞 / 組織スライス / 生体イメージング / 免疫細胞 / リンパ節 |
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| Outline of Final Research Achievements |
In order to clarify the spatiotemporal cell-cell interactions in lymph nodes, which play a fundamental role in immune responses, we prepared tissue slices from mouse lymph nodes and explored an efficient method to collect cell populations in microscopic areas. We established a method to collect small tissue fragments by covering the top and bottom of the tissue slices with a thin layer of resin film and punching them out in a circular shape using a microbiopsy trepan under a stereomicroscope. The microtissue fragments were digested with collagenase and DNase, and cells were successfully isolated. Flow cytometric analysis of the cells after staining with fluorescent antibodies confirmed that the ratio of immune cell subsets was as expected in different tissue regions.
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| Free Research Field |
免疫学
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| Academic Significance and Societal Importance of the Research Achievements |
近年、臓器の微細構造に関する理解は進んできているが、多くは固定した薄切標本を用いた観察や、観察可能な範囲に限定した一部の生体観察に基づく。一方、構造の空間的な位置情報を保ったまま生きた細胞を回収することは未だ容易ではない。また、免疫細胞の移動や分子過程が明らかになる一方、臓器ごとに免疫系の性質や挙動は大きく異なり、組織の場や局所環境を理解することが重要である。特にリンパ器官の動的・変動的な性状はそれを阻んでいる。本研究で確立された手法は、ダイナミックな臓器の生理的機能の解析とともに、幅広い臓器にも適応可能で、構造や機能、疾患に伴う病理的な組織変容を明らかにするための有用な方法となる。
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