Effects of three-dimensional culturing in a fibrous matrix on cell cycle, apoptosis, and MAb production by hybridoma cells.
The effects of culturing hybridoma cells in a three-dimensional (3-D) poly(ethylene terephthalate) (PET) fibrous matrix on cell cycle, apoptosis, metabolism, and monoclonal antibody (MAb) production were evaluated by comparing with two-dimensional (2-D) culturing on microcarrier and multiwell plate surfaces. The percentage of cells in the G1/G0 phase increased during the long-term culturing period of approximately 4 weeks. Compared to the 2-D culture systems, cells grown in 3-D matrices had higher MAb productivity for long-term culture. Decreasing serum content in the culture medium increased both MAb productivity and apoptosis. However, the 3-D culture had a greater increase in MAb productivity and a much lower apoptotic rate than the 2-D culture, especially at 0% serum. Most cells in the 3-D fibrous matrix formed large aggregates and were smaller than cells grown on a 2-D surface or in suspension. The smaller cell size allowed cells to survive better in the high-cell-density environment. The fibrous matrix also selectively retained healthy, nonapoptotic cells. These results suggested that the 3-D fibrous matrix contributed to growth arrest, protected cells to better resist low-serum environments, and reduced apoptosis, all of which contributed to the high viable cell density and volumetric MAb productivity in the long-term 3-D culture.
Luo
Jun
J
Department of Chemical Engineering, The Ohio State University, 140 West 19th Avenue, Columbus, Ohio 43210, USA.
Yang
Shang-Tian
ST
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D017209:Apoptosis / Q000502:physiology; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002453:Cell Cycle / Q000502:physiology; D002460:Cell Line; D006825:Hybridomas / Q000166:cytology / Q000378:metabolism*; D008567:Membranes, Artificial; D051379:Mice; D011093:Polyethylene Terephthalates
D000911:Antibodies, Monoclonal; D008567:Membranes, Artificial; D011093:Polyethylene Terephthalates