本文選題：PK-細(xì)胞 + 低血清培養(yǎng)基��； 參考：《動(dòng)物醫(yī)學(xué)進(jìn)展》2016年09期

【摘要】：依次采用含60、30、20、10mL/L血清濃度的低血清培養(yǎng)基馴化PK-15細(xì)胞,并給馴化好的細(xì)胞上接種豬圓環(huán)病毒2型(PCV-2),以確定PCV-2在該培養(yǎng)體系下的生長情況。結(jié)果用含60、30、20mL/L血清濃度的低血清培養(yǎng)基各進(jìn)行3代次馴化,PK-15細(xì)胞能完全適應(yīng)且生長狀態(tài)良好;當(dāng)血清濃度降至10mL/L時(shí),傳代1次細(xì)胞無法保持良好狀態(tài),細(xì)胞出現(xiàn)貼壁較差、生長停滯等現(xiàn)象。各血清濃度培養(yǎng)體系中細(xì)胞生長曲線測定結(jié)果顯示,在細(xì)胞培養(yǎng)的0、24、48、72、96h各組細(xì)胞密度與常規(guī)培養(yǎng)的對(duì)照組差別不明顯。因此用該低血清培養(yǎng)體系培養(yǎng)PK-15細(xì)胞時(shí),血清最低添加量為20mL/L。用該體系培養(yǎng)的PK-15細(xì)胞接種PCV-2后,通過熒光抗體染色測定病毒滴度。結(jié)果顯示,低血清培養(yǎng)的PCV-2病毒滴度為10~(6.5)TCID_(50)/mL,與常規(guī)條件培養(yǎng)的PCV-2對(duì)照(病毒滴度為10~(6.375 )TCID_(50)/mL)差別不明顯,表明該體系可用于PCV-2的增殖。表明研究建立了PK-15細(xì)胞低血清培養(yǎng)PCV-2體系,為PCV-2的相關(guān)研究奠定了基礎(chǔ)。
[Abstract]:The PK-15 cells were acclimated on a low serum medium containing 60 ~ 30 ~ 20ml / L serum concentration and inoculated with porcine circovirus type 2 PCV-2 to determine the growth of PCV-2 in this culture system. Results the PK-15 cells were acclimated completely and grew well when the serum concentration of PK-15 was reduced to 10mL/L, and the cells could not maintain a good condition when the serum concentration was reduced to 10mL/L, and the cells had poor adherence to the wall, and the cells could not be acclimated to the medium with low serum concentration of 60 ~ 30 ~ 20 mL / L for 3 passages. Stagnation of growth, etc. The results of cell growth curve in each serum concentration culture system showed that there was no significant difference in cell density between the control group and the normal culture group at 7296 h after cell culture. Therefore, when PK-15 cells were cultured in this low serum culture system, the minimum serum addition was 20ml / L. The PK-15 cells cultured in this system were inoculated with PCV-2 and the virus titers were detected by fluorescent antibody staining. The results showed that the titer of PCV-2 virus cultured in low serum was 10 ~ (6. 5) TCID / mL, which was not significantly different from that of PCV-2 cultured in normal condition (the virus titer was 10 ~ 6. 375 / mL), which indicated that the system could be used for the proliferation of PCV-2. The results showed that the PCV-2 system of PK-15 cells cultured with low serum was established, which laid a foundation for the study of PCV-2.
【作者單位】： 吉林大學(xué)動(dòng)物醫(yī)學(xué)學(xué)院;山東綠都生物科技有限公司;
【基金】：山東省重點(diǎn)研發(fā)計(jì)劃項(xiàng)目(2015GSF121027)
【分類號(hào)】：S852.651

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