| 细胞 | 来源 | 用途 |
| Sf9 | 从草地贪夜蛾蛹(Spodoptera frugiperda)的卵巢组织所得细胞系IPLB-SF21-AE分离得到的克隆 | 适于转染、生产高滴度病毒以及表达重组蛋白，使用最广泛 |
| Sf21 | 从草地贪夜蛾蛹(Spodoptera frugiperda)的卵巢组织所得细胞系IPLB-SF21-AE衍生所得的克隆 | 适于转染、生产高滴度病毒以及表达重组蛋白。Sf21细胞可能比Sf9细胞表达蛋白量更高。 |
| H5 | 来源于粉纹夜蛾(Trichoplusia ni)的成熟卵巢组织 | 转染效率偏低，不建议作为bacmid的转染细胞系；但是一旦得到重组病毒，HighFive细胞表达重组病毒性能优越，更适合表达分泌型重组蛋白 |
昆虫细胞在悬浮培养过程中活率低于90%，可排查以下原因：
1. 叶轮旋转剪切力的影响。可加入Pluronic F-68，使得终浓度达到所需0.1%。
2. 通风不良。①培养液体积不宜超过摇瓶容积的1/2； ②培养基体积接近最小要求体积；③叶轮旋转流畅。急动颠簸则通风不良。
3. 杆状病毒或真菌污染。①换掉旧培养基然后使用新鲜培养基。②培养基加抗生素。
4. 野毒或重组杆状病毒污染。可见细胞裂解、碎片和/或细胞鼓胀。核内容物表明野生型感染。①换掉旧培养基然后使用新鲜培养基。②在能触及杆状病毒的地方要求使用完全无菌的摇瓶。
Figure 1.
a) Passage culture of Sf9 cells using JSBio media and control medium, seeded at 1.0 × 106 cells/mL; the cell density reaches 7.0–8.0 × 106 cells/mL after 72 h.
b) Batch culture of Sf9 cells using JSBio media and control medium, seeded at 1.0 × 106 cells/mL; the peak viable cell density reaches 15.0–16.0 × 106 cells/mL.
Figure 2.
a) Passage culture of H5 cells using JSBio media and control medium, seeded at 1.0 × 106cells/mL; the cell density reaches 6.0–7.0 × 106cells/mL after 48 h.
b) Batch culture of H5 cells using JSBio media and control medium, seeded at 1.0 × 106 cells/mL; the peak viable cell density reaches 13.0–15.0 × 106cells/mL.
Figure 3.
a) Infection of Sf9 cells with viral seeds in different media; the proportion of eGFP-expressing cells was detected after infection, showing an advantage over competitor media.
b) Infection of H5 cells with viral seeds in different media; the proportion of eGFP-expressing cells was detected after infection, showing an advantage over competitor media.
Figure 4. Superior viral seed titers in Sf9 cells cultured with JSBio’s media compared to that of competitor media
Figure 5. Superior protein expression level of classical swine fever E2 in H5 cells cultured with JSBio’s media compared to that of competitor media