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首页> 外文会议>General Meeting of European Society of Animal Cell Technology >Comparison of the Cultivation of Wild and Transfected Drosophila Melanogaster S2 Cells in Different Media
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Comparison of the Cultivation of Wild and Transfected Drosophila Melanogaster S2 Cells in Different Media

机译：不同媒体中野生转染果蝇S2细胞培养的比较

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Insect cells have been increasingly employed for the production of recombinant proteins. One of the most widely used dipteran cells in transfection studies is the Schneider 2 (S2) cell line, established from Drosophila melanogaster embryonic tissue. In this work, the growth and proliferation of wild and transfected S2 cells expressing the G glycoprotein from rabies virus (GPV) were compared employing different culture media. For the transfected (S2AcGPV2) cell contruction, the plasmid pGPV encoding the sequence of interest and the vectors pAc 5.1/V5-His A under the control of the Drosophila actin promoter were utilized. The selection vector pCoHygro carrying genes coding for hygromicin-inactivating enzymes was also employed and the cells were transfected using lipofectin. Due to wide utilization of TNM-FH and TCI00 media (both requiring supplementation with fetal bovine serum) for insect cells, their low cost and the low protein content of SF900II medium, these three media and the mixture TNM-FH-SF900II (1:1) were evaluated for the choice of a suitable media for S2 cells. The results indicated that the wild and the transfected cells presented different growth characteristics in the distinct media. In SF900II medjum, larger accumulation and consumption of lactate were observed for the wild cell culture. In TC100, however, S2AcGPV2 cells did not produce lactate. In TNM-FH, the S2 cells presented lower growth rate (mu_(max)=0.0078 h~(-1)) when compared to the other media (mu_(max)=0.0375 h~(-1) andmu_(max)=0.0112 h~(-1) for SF900II and TC100, respectively), with an accentuated viability drop during the first days of culture. The mixture of TNM-FH and SF900II at a 1:1 volume ratio resulted in cell growth (mu_(max)=0.0377 h~(-1)) similar to that observed in SF900II medium only, allowing significant culture media cost reduction. Transfected cells could not be adapted to TNM-FH medium. Cell growth of S2AcGPV2 cells in TC100 medium (mu_(max)=0.0151 h~(-1)) was lower than in SF900II mediurr (mu_(max)=0.0407 h~(-1)).
机译：昆虫细胞越来越多地用于生产重组蛋白质。转染研究中最广泛使用的Dipteran细胞之一是施耐德2（S2）细胞系，从果蝇黑素转酯胚胎组织建立。在这项工作中，比较使用不同培养基的狂犬病病毒（GPV）与狂犬病病毒（GPV）的野生和转染S2细胞的生长和增殖。对于转染的（S2acGPv2）细胞标记，使用编码感兴趣的序列和在果蝇肌动蛋白启动子的控制下编码感兴趣序列和载体Pac 5.1 / V5-His A的质粒pGPV。还采用选择载体携带编码用于卫生蛋白灭活酶的基因，使用唇素转染细胞。由于TNM-FH和TCI00培养基的广泛利用（需要补充胎牛血清），用于昆虫细胞，其低成本和SF900II培养基的低蛋白质含量，这三种培养基和混合物TNM-FH-SF900II（1： 1）评估S2细胞的合适培养基。结果表明，野生和转染细胞在不同介质中呈现了不同的生长特征。在SF900II中，观察到野生细胞培养物的较大积聚和乳酸的消耗。然而，在TC100中，S2acGPv2细胞没有产生乳酸。在TNM-FH中，与其他媒体相比，S2细胞呈下较低的生长速率（MU_（MU_（MAX）= 0.0078小时〜（-1））（mu_（mu_（mu_（mu_（mu_）= 0.0375 h〜（-1）和mu_（max）= SF900II和TC100分别为0.0112 H〜（-1），在文化的第一天内，突出的活力下降。 TNM-FH和SF900II的混合物在1：1体积比下导致细胞生长（mu_（mu_（mu_（mu_）= 0.0377 h〜（-1）），其类似于在SF900II培养基中观察到的，允许显着的培养介质成本降低。转染细胞不能适应TNM-FH培养基。在TC100培养基中S2acGPv2细胞的细胞生长（MU_（MU_（MU_（MU_（MU_（MU_）= 0.0151H〜（-1））低于SF900II mediuRR（MU_（MU_（MAX）= 0.0407H〜（-1））。

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