• Journal of Life Science
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• v.24 no.11
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• pp.1252-1257
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• 2014

While dimethyl sulfoxide (DMSO) is the most commonly used cryoprotectant agent in the cryopreservation of cultured mammalian cells, it has been reported to cause differentiation of some cell lines by DNA methylation and associated histone modifications. To avoid the side effects of DMSO in cryopreservation, other agents might be more appropriate for maintaining the stable differentiation of cultured cell phenotypes through cryopreservation. All cryoprotectants should be highly soluble in water and display low cell toxicity. Cryoprotective agents have been shown to be effective in animal sperm preservation, and eight types of amides were examined in the cryopreservation of cultured mouse endothelial cells. Among the amides examined, acetamide and lactamide were effective cryoprotectants for cultured mammalian cells. The most effective concentration of lactamide, 1.5 M, had an even lower cryoprotective ability than 1M DMSO. Because successful cryopreservation of cultured cells is hampered by osmotic stress, the adequate ionic concentration was determined by diluting phosphate-buffered saline (PBS) in the 1.5M lactamide solution. The most effective concentration was $0.4{\times}PBS$, which minimized osmotic stress during the cryopreservation of cultured cells. As the addition of high molecular weight materials in cryopreservation media improves the viability of cells, the effects of bovine serum albumin (BSA), hydroxyethyl-starch (HES), and dextran were examined. The best combination of lactamide-based media for cryopreservation was found to be 1.5 M lactamide in $0.4{\times}PBS$ with 1% BSA.

• Archives of Pharmacal Research
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• v.20 no.5
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• pp.496-500
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• 1997

A strong antioxidant activity, which was measured by the radical scavenging effect on 1, 1-diphenyl-2-picrylhydrazyl(DPPH) radical, was detected in the methanol extract of Salvia miltiorrhiza Bunge (Labiatae). By activity-directed fractionation, compounds 1 and 2 were isolated as antioxidant principles of S. miltiorrhiza. Compounds 1 and 2 were identified as dimethyl lithospermate and 3-(3, 4-dihydroxyphenyl)lactamide, respectively, on the basis of spectral data. The radical scavenging effect of compounds 1 and 2 on DPPH radical exceeded that of L-ascorbic acid which is a well known antioxidant. These two compounds also showed prominent inhibitory activity against free radical generation in dichlorofluorescein (DCF) method and cytoprotective effect against t-BHP in cultured liver cell.