UMR BOREA (Biologie des Organismes et Ecosystèmes Aquatiques) MNHN/CNRS-7208/IRD-207/UPMC, Muséum national d'Histoire naturelle, Station de Biologie Marine, 29900, Concarneau, France.
Cytotechnology. 2013 Oct;65(5):737-47. doi: 10.1007/s10616-013-9576-0. Epub 2013 Aug 9.
Mollusc shell biomineralisation involves a variety of organic macromolecules (matrix proteins and enzymes) that control calcium carbonate (CaCO3) deposition, growth of crystals, the selection of polymorph, and the microstructure of the shell. Since the mantle and the hemocytes play an important role in the control of shell formation, primary cell cultures have been developed to study the expression of three biomineralisation genes recently identified in the abalone Haliotis tuberculata: a matrix protein, Lustrin A, and two carbonic anhydrase enzymes. Mantle cells and hemocytes were successfully maintained in primary cultures and were evaluated for their viability and proliferation over time using a semi-automated assay (XTT). PCR and densitometric analysis were used to semi-quantify the gene expression and compare the level of expression in native tissues and cultured cells. The results demonstrated that the three genes of interest were being expressed in abalone tissues, with expression highest in the mantle and much lower in the hemocytes and the gills. Biomineralisation genes were also expressed significantly in mantle cells, confirming that primary cultures of target tissues are suitable models for in vitro investigation of matrix protein secretion.
软体动物壳的生物矿化涉及多种有机大分子（基质蛋白和酶），它们控制着碳酸钙（CaCO3）的沉积、晶体的生长、多晶型的选择和壳的微观结构。由于套膜和血细胞在壳形成的控制中起着重要作用，因此已经开发了原代细胞培养来研究最近在鲍鱼 Haliotis tuberculata 中鉴定的三种生物矿化基因的表达：基质蛋白 Lustrin A 和两种碳酸酐酶酶。套膜细胞和血细胞成功地在原代培养中维持，并使用半自动测定法（XTT）评估其随时间的活力和增殖。PCR 和密度测定分析用于半定量基因表达，并比较天然组织和培养细胞中的表达水平。结果表明，感兴趣的三个基因在鲍鱼组织中表达，其中套膜中的表达最高，血细胞和鳃中的表达较低。生物矿化基因也在套膜细胞中显著表达，证实目标组织的原代培养是体外研究基质蛋白分泌的合适模型。
