Ligand accumulation in autocrine cell cultures.
Cell-culture assays are routinely used to analyze autocrine signaling systems, but quantitative experiments are rarely possible. To enable the quantitative design and analysis of experiments with autocrine cells, we develop a biophysical theory of ligand accumulation in cell-culture assays. Our theory predicts the ligand concentration as a function of time and measurable parameters of autocrine cells and cell-culture experiments. The key step of our analysis is the derivation of the survival probability of a single ligand released from the surface of an autocrine cell. An expression for this probability is derived using the boundary homogenization approach and tested by stochastic simulations. We use this expression in the integral balance equations, from which we find the Laplace transform of the ligand concentration. We demonstrate how the theory works by analyzing the autocrine epidermal growth factor receptor system and discuss the extension of our methods to other experiments with cultured autocrine cells.
Monine
Michael I
MI
Department of Chemical Engineering and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey 08544, USA.
Berezhkovskii
Alexander M
AM
Joslin
Elizabeth J
EJ
Wiley
H Steven
HS
Lauffenburger
Douglas A
DA
Shvartsman
Stanislav Y
SY
D019898:Autocrine Communication; D001692:Biological Transport; D001703:Biophysics / Q000379:methods; D001940:Breast / Q000166:cytology; D018929:Cell Culture Techniques; D002478:Cells, Cultured / Q000166:cytology*; D003470:Culture Media / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D004815:Epidermal Growth Factor / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology; D066246:ErbB Receptors / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008024:Ligands; D008954:Models, Biological; D008962:Models, Theoretical; D009569:Nitric Oxide / Q000737:chemistry; D010829:Physiological Phenomena; D013997:Time Factors
D003470:Culture Media; D008024:Ligands; D009569:Nitric Oxide; D004815:Epidermal Growth Factor; D066246:ErbB Receptors