冷冻保存胎鼠生物功能下颌下腺原基。
Cryopreservation of Biologically Functional Submandibular Gland Rudiments from Fetal Mice.
机构信息
Department of Oral Maxillofacial Surgery, Asahi University School of Dentistry, Gifu, Japan.
Department of Pharmacology, Asahi University School of Dentistry, Gifu, Japan.
出版信息
In Vivo. 2020 Nov-Dec;34(6):3271-3277. doi: 10.21873/invivo.12164.
BACKGROUND/AIM: Cryopreservation of cell lines has been widely used in the laboratory; however, cryopreservation of organs is still considered to be difficult. The submandibular gland (SMG) of fetal mice is one of the best-characterized organs. We investigated the conditions for cryopreserving SMG rudiments.
MATERIALS AND METHODS
Embryonic day 13 SMG rudiments were cryopreserved with or without a cryoprotectant. They were thawed and incubated in DMEM/F12 medium. Moreover, the influence of EGF stimulation on the signaling cascade after frozen-thawing the rudiments was analyzed by Western blotting.
RESULTS
When SMG rudiments were cryopreserved without a cryoprotectant, all cells in the rudiments died. However, the SMG rudiments that had been preserved in a cryoprotectant showed branching morphogenesis. Additionally, the responsiveness of signaling cascades to EGF did not differ between frozen with a cryoprotectant and non-frozen rudiments.
CONCLUSION
Cryopreservation might be a useful technology for preserving tissues from small organs, such as fetal SMG rudiments.
背景/目的：细胞系的冷冻保存已在实验室中得到广泛应用，但器官的冷冻保存仍被认为较为困难。胎儿小鼠的颌下腺（SMG）是研究得最为透彻的器官之一。我们研究了冷冻保存 SMG 原基的条件。
材料和方法
对胚胎第 13 天的 SMG 原基进行有或无冷冻保护剂的冷冻保存。解冻后，将它们在 DMEM/F12 培养基中孵育。此外，通过 Western blot 分析了 EGF 刺激对冷冻-解冻后原基信号级联的影响。
结果
当 SMG 原基未经冷冻保护剂冷冻保存时，原基中的所有细胞均死亡。然而，经冷冻保护剂保存的 SMG 原基表现出分支形态发生。此外，冷冻保护剂冷冻保存和未冷冻保存的原基对 EGF 的信号级联的反应性没有差异。
结论
冷冻保存可能是保存小型器官组织（如胎儿 SMG 原基）的一种有用技术。
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