福寿螺组织细胞原代培养及染色体核型分析

    Primary histocyte culture and chromosome karyotype analysis of Pomacea canaliculata

    • 摘要: 无菌分离取得福寿螺肌肉、外套膜、肾脏及脑组织,分别采用胰酶消化法和研磨法进行组织细胞分离,选取DMEM、F12 和改良1640 培养基分别进行细胞原代培养,光镜下观察细胞的生长状态,MTT法测定细胞活性。细胞培养一定时间后进行染色体制备及染色体核型分析。结果表明,福寿螺组织细胞在不同的培养基培养下活力具有明显差异,用含水解乳蛋白的DMEM 培养基培养的福寿螺外套膜和斧足组织细胞活力较好;用M199 培养基培养的福寿螺斧外套膜和脑组织细胞活力较好。核型分析结果表明福寿螺染色体为二倍体(2n = 28,NF = 56),核型为2B 型,核型不对称系数为66.4%。

       

      Abstract: Pomacea canaliculata’s muscle,mantle,kidney and brain tissues were aseptic separated by trypsin digestion and grinding method. Cells were primary cultured with DMEM,F12 and modified 1640 medium. The growing status was observed under light microscope. The cell viability was determined by MTT method. After a certain time,chromosome preparation and chromosome karyotype analysis were conducted. Results showed that there was significant difference in histocytes viability among different culture medium. The better viability of mantle and muscle histocytes was cultured with DMEM medium containing hydrolyzed milk protein. The better viability of mantle and brain histocytes was cultured with M199 medium. In karyotype analysis,results showed that P. canaliculata was diploid( 2n = 28,NF = 56). The nuclear type was 2B and the nuclear asymmetry coefficient was 66.4%

       

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