Department of Cryobiology and Germplasm of Animals from Kazakhstan, Institute of Zoology of Republic of Kazakhstan, 93 al-Farabi Str., Almaty 050060; and Cryoprotect LLC, Innovative Center Skolkovo, Merzlotnaya str., 29-43, Yakutsk 677010, Russia.
Cryo Letters. 2024 Mar-Apr;45(2):106-113.
Cold hardiness of insects from extremely cold regions is based on a principle of natural cryoprotection, which is associated with physiological mechanisms provided by cryoprotectants.
Since arctic cold-hardy insects are producers of highly effective cryoprotectants, in this study, the hemolymph of Aporia crataegi L. and Upis ceramboides L. from an extremely cold area (Yakutia) was tested as a secondary component of cryoprotective agents (CPA) for cryopreservation (-80 degree C) of human peripheral blood lymphocytes and skin fibroblasts.
Lymphocytes and skin fibroblasts were treated with various combinations of DMSO and hemolymph extract and step-wise cooled to -80 degree C. Post-cryopreservation cell viability was assessed by vital staining and morphological appearance.
Viability was higher when cells were frozen with a mixture containing DMSO and Upis ceramboides hemolymph compared to the cells frozen in DMSO, while cells frozen with DMSO and Aporia crataegi hemolymph did not survive. The fact that hemolymph of not every cold-resistant insect can be used as a secondary agent along with DMSO indicates that only a unique combination of hemolymph components and its compatibility with cells might result in a positive effect.
Although the use of insect hemolymph as a complementary agent in applied cryopreservation is a problem in terms of practical application, such studies could initiate new trends in the search for the most successful hemolymph-like cryoprotectant systems. https://doi.org/10.54680/fr24210110712.
来自极寒地区的昆虫的耐寒性基于自然抗冻保护原理，这与抗冻剂提供的生理机制有关。
由于北极耐寒昆虫是高效抗冻剂的生产者，因此在这项研究中，来自极寒地区（雅库特）的山荆子和乌苏里拟步甲的血淋巴被测试作为抗冻保护剂（CPA）的次要成分，用于保存（-80°C）人类外周血淋巴细胞和皮肤成纤维细胞。
用不同比例的 DMSO 和血淋巴提取物处理淋巴细胞和皮肤成纤维细胞，并逐步冷却至-80°C。通过活细胞染色和形态学外观评估冷冻保存后的细胞活力。
与单独使用 DMSO 冷冻的细胞相比，当细胞用含有 DMSO 和乌苏里拟步甲血淋巴的混合物冷冻时，细胞活力更高，而用 DMSO 和山荆子血淋巴冷冻的细胞则无法存活。并非所有耐寒昆虫的血淋巴都可以用作 DMSO 的辅助剂，这一事实表明，只有血淋巴成分的独特组合及其与细胞的相容性才能产生积极的效果。
尽管在实际应用中，将昆虫血淋巴用作补充剂在应用冷冻保存方面存在问题，但此类研究可能会引发寻找最成功的血淋巴样抗冻保护系统的新趋势。https://doi.org/10.54680/fr24210110712.
