Abstract

Background and Objectives

Providing fetal calf serum (FCS) alternatives as cell culture supplements is an important field of research to compensate for the FCS supply shortage.This study focused on preparation of fetal calf serum alternatives and their effects on growth and secretion of hybridoma cell lines.

Materials and Methods

Outdated human platelet units undergo extraction for its growth factors to be obtained. Human AB blood group plasma was also converted to serum and its growth effect was compared to FCS, hypoxanthine-thymidine (HT) and RPMI1640 as cell culture media and supplement. Cell growth indices were preliminary counting of cells , confluency as surface area of plates filled with cells, and titration of monoclonal anti-A and anti-B blood group antibodies collected from cultured mouse hybridoma cells. Statistical analysis including one sample t-test, logarithmic multiple regression curve fit, and factor analysis was done by SPSS v12 software.

Results

The four nutritional supplements of (1) human serum AB (AB), (2) human platelet extract (PLT), (3) equal mixture of AB & PLT (ABP), and (4) fetal calf serum as cell culture were examined on mouse hybridoma anti-A and anti-B monoclonal antibody producer cell lines for cell growth indices and compared with the same indices on RPMI1640 media. The growth-stimulating effects in descending order of values were (1) ABP5% , (2) FCS10% , (3) ABP10% , (4) AB5% , (5) AB10% , (6) PLT5% , (7) ABP20% , (8) PLT10% , (9) PLT20%, and (10)HT but AB20% inhibited growth of mentioned hybridoma cell lines. The titer of anti-A and anti-B monoclonal antibodies produced by cultured hybridoma on 5 and 10 percent concentration of AB , PLT and ABP compared to FCS5-10% at descending order were (1) PLT5%, (2) PLT10% , ABP5% , ABP10% , AB10%, and (3) AB5%.

Conclusions

In general FCS had the following effects on curves of cell growth: (1) the highest increase on slope of multiplication (ascending) phase, (2) the highest increase on slope of death (descending) phase, and (3) the lowest duration of stationary phase. Then, FCS can be appropriate for growth of cells at initial low cell count. Human serum AB, human platelet extract, and equal mixture of both at optimum concentrations (these supplements at high concentrations killed cells) compared to FCS showed (1) decreased slope of multiplication phase, (2) decreased slope of death phase, and (3) increased duration of stationary phase. Thus, AB and PLT may be suitable for continuous cell culture systems in which cell survival during longer times is required. Factor analysis was introduced as a model to evaluate kinetics of cell growth at different supplements.

Key words : Human serum, Cell culture, Growth kinetic, Blood groups, Factor analysis