Lang A B, Schürch U, Zimmermann F, Bruderer U
Department of Immunology, Swiss Serum and Vaccine Institute, Berne.
J Immunol Methods. 1992 Sep 18;154(1):21-6. doi: 10.1016/0022-1759(92)90208-b.
The direct propagation of newly formed human hybridomas in serum-free medium selects for hybrids with a metabolism best suited to growth in this environment. Under optimal culture conditions, this procedure results in the generation of antigen-specific human hybridomas comparable in frequency, stability, and antibody secretion rate to that obtained with murine hybridomas. After a transient phase of a few days in the appropriate selection medium supplemented with 1% serum, hybridomas grow in serum-free medium in stationary cultures with a cell doubling time of 15-25 h and an antibody production rate averaging 12 micrograms/10(6) cells/day. Clones propagated in bioreactors exhibited a cell doubling time of 29-35 h and an antibody secretion rate of 10-21 micrograms/10(6) cells/day.
新形成的人杂交瘤在无血清培养基中的直接传代选择出了代谢最适合在这种环境中生长的杂交瘤。在最佳培养条件下,该程序导致产生抗原特异性人杂交瘤,其频率、稳定性和抗体分泌率与鼠杂交瘤相当。在补充有1%血清的合适选择培养基中培养几天后的短暂阶段后,杂交瘤在无血清培养基的静止培养物中生长,细胞倍增时间为15 - 25小时,抗体产生率平均为12微克/10⁶细胞/天。在生物反应器中传代的克隆细胞倍增时间为29 - 35小时,抗体分泌率为10 - 21微克/10⁶细胞/天。