泊洛沙姆188、碳酸氢钠和HEPES对CPV ID3细胞株生长的影响  

Effects of Poloxamer 188,Sodium Bicarbonate and HEPES on the Growth of CPV ID3 Cell

作　　者：孙晓莹 李国军 王延涛 侯喜林[1] Sun Xiaoying;Li Guojun;Wang Yantao;Hou Xilin(College of Animal Science and Technology,Heilongjiang Bayi Agricultural University,Daqing 163319;Shanghai Maibang Biological Technology Co.,Ltd.)

机构地区：[1]黑龙江八一农垦大学动物科技学院,大庆163319 [2]上海迈邦生物科技有限公司

出　　处：《黑龙江八一农垦大学学报》2022年第2期32-37,43,共7页journal of heilongjiang bayi agricultural university

摘　　要：为了研究不同浓度的泊洛沙姆188(Poloxamer 188,P188)、碳酸氢钠(NaHCO_(3))和HEPES对CPV ID3细胞株生长的影响,分别向无血清培养基中添加不同浓度的P188、NaHCO_(3)和HEPES于125 mL摇瓶内悬浮培养稳定表达犬细小病毒单抗的CPV ID3细胞株,培养体积30 mL,培养条件为36.5℃,5%CO_(2),120 rpm,每天取样测量细胞的密度、活率,并于培养结束后检测抗体表达量。结果显示,P188在0~15 g·L^(-1)的添加范围内,P188浓度越高,细胞高密度维持时间越长,抗体表达量越高。NaHCO_(3)在1.5~2.5 g·L^(-1)添加范围内,NaHCO_(3)浓度与细胞生长速率,乳酸积累量成正比,与抗体表达量成反比。HEPES在0~25 mM添加范围内,与抗体表达量成正比,与最高活细胞密度成反比,当HEPES浓度为10 mM时,细胞生长速率最大;当HEPES浓度大于10 mM时,HEPES浓度与细胞生长速率成反比。结论:针对CPV ID3细胞株,在无血清培养基中添加P188可以延长细胞密度维持时间,提高抗体表达量;培养基中的NaHCO_(3)和HEPES的最适浓度分别为2 g·L^(-1)和10 mM。In order to study the effects of different concentrations of Poloxamer 188(P188),sodium bicarbonate(NaHCO_(3))and HEPES on the growth of CPV ID3 cell line,different concentrations of P188,NaHCO_(3) and HEPES were added to the serum-free medium in 125 mL.Suspension culture of CPV ID3 cell line stably expressing canine parvovirus monoclonal antibody in a flask,with a culture volume of 30 mL,culture conditions at 36.5℃,5%CO_(2),and 120 rpm.Samples were taken every day to measure cell density and viability,And detected the production of antibody after the end of the culture.The results showed that P188 was in the range of 0~15 g·L^(-1),the higher the P188 concentration,the longer the cell density was maintained,and the higher the antibody expression.When NaHCO_(3) was added in the range of 1.5~2.5 g·L^(-1),the concentration of NaHCO_(3) was directly proportional to the specific cell growth rate and lactic acid accumulation,and inversely proportional to the antibody expression.When HEPES was added in the range of 0~25 mM,it was directly proportional to the antibody expression and inversely proportional to the highest viable cell density.When the concentration of HEPES was 10 mM,the cell growth rate was the largest;when the concentration of HEPES is greater than 10 mM,the concentration of HEPES is inversely proportional to the cell growth rate.Conclusion:For CPV ID3 cell line,adding P188 to the serum-free medium used in this study can prolong cell density maintenance time and increase antibody expression;the optimal concentrations of NaHCO_(3) and HEPES in the medium are 2 g·L^(-1)and 10 mM.

关 键 词：泊洛沙姆188 碳酸氢钠 HEPES 无血清培养基 杂交瘤细胞 

分 类 号：Q813.1[生物学—生物工程]