ARTP诱变对三角帆蚌外套膜细胞生长活性及其生物矿化相关因子的影响

为获取高活力的外套膜细胞,研究通过常压室温等离子体(Atmospheric and room temperature plasma,ARTP)诱变和流式细胞术等技术分析了不同诱变气量组(10、12和15 SLM组)和处理时间对三角帆蚌(Hyriopsis cumingii)体外培养的外套膜细胞的细胞活性及生物矿化相关功...

EFFECTS OF MUTAGENESIS BY ATMOSPHERIC AND ROOM TEMPERATURE PLASMA(ARTP)ON THE GROWTH AND BIOMINERALIZATION RELATED ACTIVITY OF THE MANTLE CELL MUTAGENESIS IN HYRIOPSIS CUMINGII

The mantle of pearl oyster is one of the indispensable tissues for pearl cultivation.In order to obtain highactivity mantle cells,the effects of different gas volume groups10,12 and 15 SLM(standard liter per minute;SLM)]and treatment time on cell activity and biomineralizated functions of mantle cells of Hyriopsis cumingii were analyzed using ARTP mutagenesis and flow cytometry.The results showed that the activity of mantle cells of Hyriopsis cumingii increased significantly in 360—900s by ARTP,and reached the maximum value at 900s(P<0.05),which was consistent with the cell activity results of osmotic stabilizer groups.Mantle cells activity in 12 and 15 SLM groups increased significantly at 360—900s(P<0.05),and the proliferation index(PI)of mantle cells reached the maximum value in 12 SLM group(P<0.05);in addition,cells were cultured in vitro for 24h after mutagenesis,the viability of mantle cells in 12 SLM group at 720s was the highest(P<0.05),and the SOD activity(mutagenesis time was 720s,the same as below)showed a significant decreasing trend with the increase of mutagenesis gas volume,and it decreased significantly to the lowest level in the 15 SLM group(P<0.05).On the contrary,the micronucleus rate reached the maximum value;biomineralization analysis showed that the concentration of Ca²⁺,the key enzymes related to biomineralization(carbonic anhydrase,alkaline phosphatase)and calmodulin(CAM)genes of mantle cells reached the maximum in the 12 SLM group(P<0.05),but expression levels of EFCB1(EF-hand calcium-binding domain-containing protein 1)gene reached the maximum in the 10 SLM group(P<0.05),followed by 12 SLM group.The above findings indicated that ARTP mutation had the most significant effect on the cell activity and other biological activity of mantle cells when the gas volume is 12 SLM for 720s and the osmotic pressure stabilizer is used together with ARTP,indicating that ARTP mutagenesis can be effective in vitro culture of mantle cells to provide a biological basis and a new idea for the establishment of cell lines.