表皮生长因子在无血清条件下促进 E6 和 CCL-81 Vero 细胞增殖

Diena Delaiah, Ahmad Aswin, Helen Susilowati, Andi Yasmin Wijaya, Firdausy Kurnia Maulana, D. Diyantoro, Watchareewan Rodprasert, Yulianna Puspitasari, Y. Dhamayanti, S. Kuncorojakti
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引用次数: 0

摘要

将 Vero 细胞培养作为生产病毒疫苗的平台,是疫苗生产中一个成熟的标准化流程。一般来说,Vero 细胞培养需要培养基作为营养源,并补充血清以提供生长因子。然而,血清有几个缺点,包括批次间的差异和偶联剂。因此,人们使用标准化的生长因子配制化学定义的无血清培养基(SFM)。表皮生长因子(EGF)是一种在无血清培养基系统中显示出充分有丝分裂支持的生长因子,尤其是在 Vero 细胞中。在本研究中,SFM-EGF 培养基与补充血清的培养基(即 MEM 10%和 MEM 5% FBS)进行了比较。细胞形态用倒置显微镜观察,细胞增殖用 MTT 比色法评估。Vero E6 和 Vero CCL-81 细胞形态未发生任何变化。第 1-4 天,Vero E6 和 Vero CCL-81 在 SFM-EGF 培养基中的增殖与在 MEM 10% 或 MEM 5% 血清补充培养基中的增殖相比没有显著差异。不过,给予 SFM-EGF 培养基的 Vero E6 和 Vero CCL-81 细胞的 OD 值平均低于 MEM 10%,但高于补充 5% FBS 的 MEM。因此,这项研究证明,使用 SFM-EGF 可以促进 Vero E6 和 Vero CCL-81 细胞的增殖。
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Epidermal Growth Factor Promotes E6 and CCL-81 Vero Cells Proliferation Under Serum-Free Condition
Vero cell culture as a platform for producing viral vaccines is an established and standardized process in vaccine manufacture. Generally, Vero cell culture requires media as a source of nutrition with serum supplementation to provide growth factors. However, the serum has several disadvantages including batch to batch variation and adventitious agent. Therefore, chemically defined serum-free media (SFM) are formulated by using standardized growth factors. Epidermal Growth Factor (EGF) is one of growth factors that showed adequate mitogenic support in serum-free medium system, especially in Vero cells. In this study, SFM-EGF media was compared with serum supplementation media, namely MEM 10% and MEM 5% FBS supplementation. The cell morphology was observed using an inverted microscope and their proliferation was evaluated using a MTT colorimetric-based assay. Vero E6 and Vero CCL-81 cells morphology did not show any morphological changes. Vero E6 and Vero CCL-81 proliferation in SFM-EGF media on day one to four did not show a significant difference compared to MEM 10% or MEM 5% serum supplementation media. However, the OD values of both Vero E6 and Vero CCL-81 cells given SFM-EGF media produced an average value below MEM 10% but higher than MEM 5% FBS supplementation. As such, this study proved that utilizing SFM-EGF could support the proliferation of Vero E6 and Vero CCL-81 cells.
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