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为得到大颗粒优质的淡水有核珍珠,进行了游离细胞植入法在三角帆蚌(Hyriopsis cumingii)内脏团插核育珠的研究。实验采用两种培养基(培养基1、2)进行外套膜外膜细胞的培养,对不同培养时间(2、4、6、12 h)的细胞活力进行检测,同时将处理过的珠核分别置于两组细胞悬液中共培养,6 h后采用内脏团插核手术,对500只三角帆蚌进行插核,并进行为期5个月的淡水有核珍珠生产实验。实验结果表明:细胞活力随培养时间逐渐增大,培养到6、12 h时,两种培养基中的细胞活力较2、4 h时均有显著提高(P<0.05),6 h与12 h相比差异不显著(P>0.05);细胞在两种培养基中分别培养,在培养2 h时两组间细胞活力没有显著差异(P>0.05),而在培养4、6、12 h时培养基2组的细胞活力显著高于培养基1组(P<0.05);培养基2组培养的细胞孵育珠核6 h后,贴附的细胞数量明显增多且分布均匀,插核5个月后,形成了包裹完整、具有光泽、沉积0.8 mm珍珠质的大型珍珠,而采用培养基1组培养的外套膜细胞进行珠核孵育而后插核,得到的素珠较多,且没有形成完整珍珠质包被的珍珠。实验表明改进后的培养基有利于三角帆蚌外套膜细胞培养,从而有利于内脏团有核珍珠的培育,这为进一步开展淡水贝类细胞培养的研究提供了实践基础,为大型有核珍珠的培育提供了依据。研究亮点:本文对体外培养三角帆蚌外套膜细胞的培养基进行改进,添加牛磺酸、Ca2+等适宜细胞生长以及珍珠质沉积的成分,促进了细胞的生长。利用改进后的游离细胞悬液与珠核共培养,并进行内脏团插核手术,5个月后得到了包被完整、具有光泽、沉积0.8 mm珍珠质的大型有核珍珠。
In order to obtain large freshwater and nucleated pearl with large particles, a free cell implantation study was conducted on visceral pellets of Hyriopsis cumingii. The experiment used two kinds of culture medium (culture medium 1 and 2) to culture the epicardial cells of the mantle and tested the cell viability at different culture time (2, 4, 6, 12 h) The cells were co-cultured in two cell suspensions. Six hours later, 500 Hyriopsis cumingii were inserted into the nucleus and the freshwater nuclear pellets were produced for 5 months. The results showed that the viability of cells increased with the time of incubation, and the viability of both media increased significantly (P <0.05) at 6 h and 12 h (P> 0.05). The cells were cultured in two mediums respectively. There was no significant difference in cell viability between the two groups at 2 h (P> 0.05), while at 4, 6 and 12 h The cell viability of medium 2 was significantly higher than that of medium 1 (P <0.05). After incubated for 6 h in medium 2, the number of attached cells was significantly increased and distributed evenly. After the formation of a complete package, luster, deposition of 0.8 mm nacre of large pearl, and culture medium using a group of cultured mantle cells incubated with beads and then insert nuclei, resulting in more beads, and did not form a complete pearl Quality coated pearls. Experiments show that the improved medium is conducive to the culture of H. brachiaria mantle cells, which is conducive to the development of visceral mass of pearl nucleus, which provides a practical basis for the further development of freshwater shellfish cell culture research for the large nuclear pearl Provide a basis for cultivation. Research highlights: In this paper, we improved the culture medium of M. mandshurica cultured in vitro, and added the proper cell growth and the components of nacre deposited by Taurine, Ca2 + to promote the growth of cells. Five months after the co-culture of the modified free cell suspension and the nucleus, the nucleated pearl with a gloss of 0.8 mm was deposited.