Koseki H, Kaku M, Kawata T, Kojima S, Sumi H, Shikata H, Motokawa M, Fujita T, Ohtani J, Tanne K
Department of Orthodontics and Craniofacial Developmental Biology, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan.
Cryo Letters. 2013 Jan-Feb;34(1):10-9.
In order to determine a suitable condition for osteoblasts cryopreservation, murine osteoblasts were freezed by programmed freezer with a magnetic field (CAS freezer). After 7 days cryopreservation at -150°, the number of survival cells immediately after thawing and the growth rate of cultured cells for 48 hours were examined. Gene and protein expression of alkaline phosphatase (ALP), osteopontin (OPN) and bone sialoprotein (BSP) were compared between cryopreserved and non-cryopreserved groups. As a result, a plunging temperature of -30°, a hold-time at -5° for 15 minutes and a 0.1 mT of magnetic field led to the largest survival and growth rate. Moreover, there was no significant difference in ALP, OPN and BSP mRNA and protein expression between cryopreserved and control groups. From these results, it was suggested that the CAS freezer is available for osteoblast cryopreservation and bone tissue banking can be established in the future.
为了确定成骨细胞冷冻保存的合适条件，将小鼠成骨细胞用带磁场的程序降温冷冻仪（CAS冷冻仪）进行冷冻。在-150°冷冻保存7天后，检测解冻后立即存活的细胞数量以及培养48小时的细胞生长率。比较冷冻保存组和未冷冻保存组碱性磷酸酶（ALP）、骨桥蛋白（OPN）和骨涎蛋白（BSP）的基因和蛋白表达。结果显示，-30°的降温速率、-5°保持15分钟以及0.1 mT的磁场导致最大的存活率和生长率。此外，冷冻保存组和对照组之间ALP、OPN和BSP的mRNA和蛋白表达没有显著差异。从这些结果表明，CAS冷冻仪可用于成骨细胞的冷冻保存，未来可以建立骨组织库。
