Abstract
The cell cycle kinetics of F3(B6) mouse hybridoma was examined by immunocytochemical staining of bromodeoxyuridine incorporated into the DNA of exponentially growing cells in three different cultures: one supplemented with 10% fetal bovine serum and two adapted to serum-free media, TABIES and BITES. The serum-free cultures, particularly the BITES, had longer cycling times and higher specific antibody production rate. Both observations were correlated to the prolongation of the G1 phase traverse time and substantiated with a starvation blocking experiment.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
AbrahamKA, EikhomTS, DowbenRM, Garatun-TjeldstøO (1976) Cell-free translation of messenger RNA for a myeloma light chain prepared from synchronised plasmacytoma cells. Eur. J. Biochem. 65: 79.
AdamG (1981) Three-state models of the cell cycle describing dependence of transition probabilities on cell density. In RotenbergM (eds) Biomathematics and Cell Kinetics. Amsterdam: Elsevier/North-Holland, p. 177.
Al-RubeaiMA, RookesS, EmeryAN (1989) Flow cytometric studies during synchronous and asynchronous suspension cultures of hybridoma cells. In: SpierRE, GriffithsJB, StephenneJ, CrooyPJ (eds.) Advances in Animal Cell Biology and Technology for Bioprocesses. London: Butterworths, p. 241.
BarnesD, SatoG (1980) Serum-free cell culture: A unifying approach. Cell 22: 649.
BasergaR (1985) The Biology of Cell Reproduction. Cambridge, Massachusetts, Harvard University Press.
DolbeareF, GratznerH, PallaviciniMG, GrayJW (1983) Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine. Proc. Natl. Acad. Sci. USA 80: 5573.
ErnstDN, McQuittyDN, WeigleWO, HobbsMV (1988) Expression of membrane activation antigens on murine B lymphocytes stimulated with lipopolysaccharide. Cell. Immunol. 114: 161.
Garatun-TjeldstøO, PrymeIF, WeltmanJK (1976) Synthesis and secretion of light-chain immunoglobulin in two successive cycles of synchronized plasmacytoma cells. J. Cell Biol. 68: 232.
GerdesJ, LemkeH, BaischH, WackerH, SchwabU, SteinH (1984) Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J. Immunol. 133: 1710.
GlassyMC, TharakanJP, ChauPC (1988) Serum-free media in hybridoma culture and monoclonal antibody production. Biotechnol. Bioeng. 32: 1015.
GrayJW (1976) Cell-cycle analysis of perturbed cell populations: computer simulation of sequential DNA distributions. Cell Tissue Kinet. 9: 499.
GuiguetM, D'HoogheMC, ValleronA-J (1980) S, G2 and M phases durations are relatively as variable as G1 duration. Cell Tissue Kinet. 13: 689.
LloydD, PooleRK, EdwardsSW (1982) The cell division cycle: temporal organization and control of cellular growth and reproduction. London, Academic Press.
MillerWM, BlanchHW, WilkeCR (1988) A kinetic analysis of hybridoma growth and metabolism in batch and continuous suspension cultures: effect of nutrient concentration, dilution rate and pH. Biotechnol. Bioeng. 32: 947.
Murakami H (1989) Serum-free media used for cultivation of hybridomas. (eds) Monoclonal Antibodies: Production and Applications. Alan R. Liss, p. 107.
MurakamiH, MasuiH, SatoG (1982) Suspension culture of hybridoma cells in serum-free medium: soybean phospholipids as the essential components. In SatoGH, PardeeAB, SirbaskuDA (eds) Growth of Cells in Hormonally Defined Media. New York: Cold Spring Harbor, p. 711.
SchliermannM, BeckersC, MiltenburgerHG (1987) Cell cycle analysis by flow cytometry in an antibody secreting cell line. Dev. Biol. Standard 66: 101.
SchneiderY-J (1989) Optimisation of hybridoma cell growth and monoclonal antibody secretion in a chemically defined, serum- and protein-free culture medium. J. Immunol. Meth. 116: 65.
SmithJA, MartinL (1973) Do cells cycle? Proc. Natl. Acad. Sci. USA 70: 1263.
TakabayashiK, SuzukiN, KannoM, ImaiK, TokuhisaT, TomiokaH, TaniguchiM (1983) Cell cycle-dependent expression of antigen-binding and I-J-bearing molecules on suppressor T cell hybridomas. J. Immunol. 130: 2552.
TharakanJP, LucasA, ChauPC (1986) Hybridoma growth and antibody secretion in serum-supplemented and low protein serum-free media. J. Immunol. Meth. 94: 225.
WatanabeS, YagiY, PressmanD (1973) Immunoglobulin production in synchronized cultures of human hematopoietic cell lines. II. Variation of synthetic and secretion activities during the cell cycle. J. Immunol. 111: 797.
WilliamsLS, NordbergJA, ChauPC (1990) Evaluation of immunocytochemical detection methods of incorporated bromodeoxyuridine in hybridomas. Cytometry 11: 490.
YanagisawaM, DolbeareF, TodorokiT, GrayJW (1985) Cell cycle analysis using numerical simulation of bivariate DNA/bromodeoxyuridine distributions. Cytometry 6: 550.
ZetterbergA, LarssonO (1985) Kinetic analysis of regulatory events in G1 leading to proliferation or quiescence of Swiss 3T3 cells. Proc. Natl. Acad. Sci. USA 82: 5365.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Richieri, R.A., Williams, L.S. & Chau, P.C. Cell cycle dependency of monoclonal antibody production in asynchronous serum-free hybridoma cultures. Cytotechnology 5, 243–254 (1991). https://doi.org/10.1007/BF00556294
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00556294