Membrane permeability coefficients of murine primary neural brain cells in the presence of cryoprotectant
Membrane permeability coefficients of murine primary neural brain cells in the presence of cryoprotectant
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DOI：
10.1016/j.cryobiol.2009.02.008
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发表时间：
2009-06-01
期刊：
影响因子：
2.7
通讯作者：
Dunnett, S. B.
中科院分区：
文献类型：
--
作者：
Paynter, S. J.;Andrews, K. J.;Dunnett, S. B.
从大脑中分离的神经细胞具有许多研究和临床应用，包括移植到患有神经退行性疾病的患者。组织供应是临床移植的主要限制因素之一。原代神经细胞的冷冻保存将改善供应，帮助组织移植手术，并促进研究。迄今为止，使用标准方法的冷冻保存已导致原代神经组织的产率和/或活力降低。为了优化专门用于这些细胞的冷冻方案，测定了非渗透体积（V-b）、水渗透性（L-p）和对冷冻保护剂的渗透性（P-cpa）。为了测定V-b，在暴露于蔗糖的非等渗溶液（150-1500 mOsmol/kg）后测量细胞体积。通过测量暴露于1.5 mol/l冷冻保护剂期间的细胞体积，测定GE细胞的L-p（μ m/min.atm）和P-cpa（μ m/s）。用电子粒子计数法测定细胞体积，Ctx和GE的V-b为27%，VM为30%。GE细胞的渗透反应在丙二醇和二甲基亚砜的存在下是相似的。在乙二醇的存在下，细胞体积的减少是更大的初始暴露于冷冻保护剂和恢复较慢。L-p的差异，而不是P-cpa，被发现之间的冷冻protectings. Present结果提供了关键参数的冷冻方案的优化冷冻保存的原代胎脑组织的神经细胞移植的应用。(C)2009爱思唯尔公司All rights reserved.
Neural cells isolated from the brain have a number of research and clinical applications, including transplantation to patients with neurodegenerative conditions. Tissue supply is one of the major limiting factors to clinical transplantation. Cryopreservation of primary neural cells would improve supply, aid in organisation of transplantation surgery and facilitate research. To date, cryopreservation using standard methods has resulted in reduced yield and/or viability of primary neural tissue. In order to optimise freezing protocols specifically for such cells, the non-osmotic volume (V-b), water permeability (L-p) and permeability to cryoprotectant (P-cpa) were determined.Murine foetal brain tissue from the ganglionic eminence (GE), ventral mesencephalon (VM), or neocortical mantle (Ctx) was trypsinised to a single cell suspension. To determine V-b, cell volume was measured after exposure to anisotonic solutions of sucrose (150-1500 mOsmol/kg). L-p (mu m/min.atm) and P-cpa (mu m/s) were determined for GE cells by measuring cell volume during exposure to 1.5 mol/l cryoprotectant. Cell volume was determined using an electronic particle counting method.V-b was 27% for Ctx and GE, and 30% for VM. The osmotic response of GE cells was similar in the presence of propane-1,2-diol and dimethyl sulphoxide. In the presence of ethylene glycol, cell volume decrease was greater on initial exposure to cryoprotectant and recovery slower. Differences in L-p, but not P-cpa, were found between cryoprotectants.The present results provide key parameters for optimisation of freezing protocols for cryopreservation of primary foetal brain tissues for application in neural cell transplantation. (C) 2009 Elsevier Inc. All rights reserved.