2021 Fiscal Year Final Research Report
Development of organ cryopreservation techniques using culture system for embryonic submandibular gland rudiments
| Project/Area Number |
19K10193
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 57040:Regenerative dentistry and dental engineering-related
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| Research Institution | Asahi University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
佐藤 慶太郎 明海大学, 歯学部, 准教授 (10549041)
設楽 彰子 朝日大学, 歯学部, 准教授 (30508718)
大野 雄太 朝日大学, 歯学部, 講師 (30796644)
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | 顎下腺原基 / 分枝形態形成 / 凍結保護薬 / 上皮成長因子 / ERK1/2 / AKT / リン酸化 |
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| Outline of Final Research Achievements |
We investigated the conditions for organ cryopreservation using the submandibular gland rudiments of embryonic mice. Cryoprotective agents were added to submandibular gland rudiments on the day 13 of gestation and cryopreserved. Some of the rudiments were also slow-frozen using Mr. Frosty to examine their effects. After thawing the submandibular gland rudiments, organ culture was performed on membranes floating in DMEM/F12 medium, and the number of branches formed was measured to examine branching morphogenesis ability. The status of phosphorylated ERK1/2 and phosphorylated AKT induced by EGF on cultured submandibular gland rudiments was also examined. The results showed that several cryoprotectants showed protection against freeze damage, and Mr. Frosty slightly enhanced the protective effect. The results suggest that cryoprotectants are effective for cryopreservation of submandibular gland primordia that retain biological activity.
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| Free Research Field |
歯科薬理学
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| Academic Significance and Societal Importance of the Research Achievements |
株化細胞の凍結保存は基礎研究分野で日常的に使用されている。しかし、生体の器官や臓器の凍結技術は確立されていない。胎生期のマウス顎下腺原基は、およそ300micrometer径の大きさの未成熟な器官で、体外へ取り出してフィルター上で器官培養することで上皮の分枝形態形成が観察できる。本研究では、株化細胞用の凍結保護薬が器官凍結に際しても保護効果を有することを明らかにした。顎下腺原基は小さい器官であり、凍結保護薬の浸透速度が適切であるため、器官凍結の保護効果を発揮すると推察された。今後さらに凍結保護薬に工夫をすることで、より大きな器官あるいは臓器を凍結保存可能であると考えられた。
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