Dessai Shanti N
Physiology and Biochemistry Laboratory, Department of Zoology, Goa University, Panaji, Goa, India.
Cryobiology. 2018 Jun;82:93-98. doi: 10.1016/j.cryobiol.2018.04.002. Epub 2018 Apr 4.
Laboratory friendly, cryopreservation procedures with respect to cryopreservation formulations and cryopreservation temperatures were attempted, in the present study to ensure perennial availability of cultured mantle cells of bivalve (Paphia malabarica). Screening of cryopreservative formulations with different concentrations of DMSO, Propylene glycol and Glycerol was carried out for cryopreservation of freshly dissociated cells of Paphia malabarica. Out of these cryopreservative formulations, 10% DMSO, 10% Propylene glycol and 15% Glycerol were selected for cryopreservation of the mantle cells pooled from 1-day old primary culture and cell line after 3 passages at the end of different cryopreservation periods. Cryopreservative formulation with 15% glycerol, served as a best cryoprotectant for the cryopreservation of cells sourced from freshly dissociated cells as well as from primary cultures and cell cultures after three passages of mantle cells of Paphia malabarica, retaining metabolic activity of resurrected cells. Both, cell cultures established from uncryopreserved cells as well as cryopreserved cells showed similar alkaline phosphatase and carbonic anhydrase activities thus indicating retention of their biomineralization capacity even after cryopreservation at low and ultralow temperatures.
在本研究中，尝试了对双壳贝类（马拉巴帕菲蛤）培养的外套膜细胞进行实验室友好型的冷冻保存程序，包括冷冻保存配方和冷冻保存温度，以确保其常年可用。对不同浓度的二甲基亚砜（DMSO）、丙二醇和甘油的冷冻保护剂配方进行了筛选，用于冷冻保存新鲜解离的马拉巴帕菲蛤细胞。在这些冷冻保护剂配方中，选择了10%DMSO、10%丙二醇和15%甘油，用于在不同冷冻保存期结束时，对来自1日龄原代培养物和传代3次后的细胞系的合并外套膜细胞进行冷冻保存。含15%甘油的冷冻保护剂配方，对来自新鲜解离细胞以及马拉巴帕菲蛤外套膜细胞传代3次后的原代培养物和细胞培养物来源的细胞冷冻保存而言，是最佳的冷冻保护剂，能保留复苏细胞的代谢活性。从未冷冻保存的细胞以及冷冻保存的细胞建立的细胞培养物，均显示出相似的碱性磷酸酶和碳酸酐酶活性，这表明即使在低温和超低温冷冻保存后，它们的生物矿化能力仍得以保留。
