杂交瘤细胞在玻璃微载体培养中生物特性观察

Biological Properties of Hybridoma Cells Grown in Glass Microcarrier Culture

本文报道以玻璃微载体小珠(CMB,Class microcarrier beads)作为载体培养杂交瘤细胞方法的建立。结果表明4株杂交瘤细胞在GMB培养显示GNB法上清中特异性单克隆抗体的滴度。OD值较单层培养法为高。比较培养7d后的杂交瘤细胞数,也以GMB法获得细胞数为多。各株细胞都诱生出高滴度腹水。亚类鉴定方面,GMB法培养的4株杂交瘤细胞上清内McAb所属亚类,又都保持了与单层法培养者完全相同的结果。表明杂交瘤细胞在CMB培养中,细胞迅速增殖,且保持各细胞株分泌其特异的McAb的生物特性,细胞活力旺盛,为今后体外培养杂交瘤细胞,制备McAb,提供了新的途径。

A method of hybridoma cell grown on the glass microcarrier beads (GMB)was developed. Four established hybridoma cell lines were continuously cultured for 7 days on the glass microcarrier beads and compared with the monolayer culture method simultaneously. The results of ELISA showed that the McAb titers and OD values of the supernatents produced from GMB culture were always higher than those obtained from the monolayer hybridoma culture at the same time. It was shown also the numbers of hybridoma cells harvested from GMB culture for 7 days were higher than those of monolayer hybridoma cells respectively. All of the 4 hybrdoma cell lines can induce mouse ascites containing specific McAbs with high titer produced by GMB culture as well as the monolayer culture. Furthermore subclasses of McAbs produced from the 4 hybridoma culture by both methods were identified by a modified ni trocellulose-ELISAtechnique It is declared that the hybridoma cells grew and multiplied well in GMB culture with high activit and kept the biological properties of secreting McAb specific to the original antigen. It will be a promising route 10 culture hybridoma cells and to produce McAb.