Modulation of antifreeze activity and the effect upon post-thaw HepG2 cell viability after cryopreservation
Modulation of antifreeze activity and the effect upon post-thaw HepG2 cell viability after cryopreservation
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DOI：
10.1016/j.cryobiol.2015.01.002
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发表时间：
2015-04-01
期刊：
影响因子：
2.7
通讯作者：
Ben, Robert N.
中科院分区：
文献类型：
--
作者：
Capicciotti, Chantelle J.;Poisson, Jessica S.;Ben, Robert N.
大多数抗冻蛋白（AFP）具有两种类型的“抗冻活性”-热滞后（TH）和冰重结晶抑制（IRI）活性。TH活性的机制已被深入研究，是AFP吸附到具有冰结合面（IBF）的冰表面的结果。相比之下，冰重结晶及其抑制的机制是相当少的理解。在本文中，我们研究了几种不同的抗冻蛋白，糖蛋白和突变体的多年生黑麦草APP（LpAFP），以了解如何IRI活动调制独立的TH活动。本研究还检查了各种AF（G）Ps保护HepG 2细胞免受冷冻损伤的能力。解冻后细胞活力与TH、IRI活性、动态冰形能力和单个冰晶生长进程相关。虽然这些结果表明AF（G）Ps作为冷冻保护剂是无效的，但它们强调了冰晶习性以及最重要的是冰生长进展如何影响冷冻保存期间的HepG 2细胞存活。(C)2015 Elsevier Inc. All rights reserved.
Most antifreeze proteins (AFPs) exhibit two types of "antifreeze activity" -thermal hysteresis (TH) and ice recrystallization inhibition (IRI) activity. The mechanism of TH activity has been studied in depth and is the result of an adsorption of AFPs to the surface of ice with an ice-binding face (IBF). In contrast, the mechanism of ice recrystallization and its inhibition is considerably less understood. In this paper, we examine several different antifreeze proteins, glycoproteins and mutants of the Lolium perenne APP (LpAFP) to understand how IRI activity is modulated independently of TH activity. This study also examines the ability of the various AF(G)Ps to protect HepG2 cells from cryoinjury. Post-thaw cell viabilities are correlated to TH, IRI activity as well as dynamic ice shaping ability and single ice crystal growth progressions. While these results demonstrate that AF(G)Ps are ineffective as cryoprotectants, they emphasize how ice crystal habit and most importantly, ice growth progression affect HepG2 cell survival during cryopreservation. (C) 2015 Elsevier Inc. All rights reserved.