Abstract
3T3-L1 preadipocytes are often being used in research of adipose-related diseases such as obesity, insulin resistance, and hyperlipidemia. We developed a stochastic model that simulated differentiation of four 3T3-L1 culture conditions distinct by the insulin concentration in the differentiation medium (2.5, 5, 7.5, or 10 μg/mL). The model simulated culture behavior and the accumulation of lipid droplets in the maturing cells from the day of induction of differentiation through 28 days after that. The cellular processes including cell adhesion, mitosis, growing after undergoing mitosis, commitment to the adipocyte lineage, and apoptosis were referred to as stochastic events in the modeling. By minimizing the error between our model and experimental results, we found that the probability for becoming committed to the adipocyte lineage in a single division and the probability for growing after undergoing mitosis were 0.02 and 0.8, respectively, regardless of the insulin concentration. The probability for undergoing mitosis was equal to 0.2 and 0.4 in cultures that had insulin concentrations of 2.5 and 5–10 μg/mL in the differentiation medium, respectively; hence the insulin concentration affected the probability for mitosis in the 3T3-L1 cells. The model and resulted probabilities now allow quantitative and visual predictions of adipogenesis in 3T3-L1 cultures, toward computational design of cell culturing protocols.
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Abbreviations
- LD:
-
Lipid droplet
- FOV:
-
Field of view
- 3D:
-
Three dimensions
References
Ailhaud, G., E. Amri, C. Cermolacce, P. Djian, C. Forest, D. Gaillard, P. Grimaldi, J. Khoo, R. Négrel, and G. Serrero-Davé. Hormonal requirements for growth and differentiation of ob17 preadipocyte cells in vitro. Diabetes Metab. 9(2):125–133, 1983.
Alber, M. S., M. A. Kiskowski, J. A. Glazier, and Y. Jiang. On cellular automaton approaches to modeling biological cells. In: Mathematical Systems Theory in Biology, Communication, and Finance, edited by J. Rosenthal, and D. S. Gilliam. New York: Springer, 2002, pp. 1–40.
Amri, E. Z., P. Grimaldi, R. Négrel, and G. Ailhaud. Adipose conversion of ob17 cells: insulin acts solely as a modulator in the expression of the differentiation program. Exp. Cell. Res. 152(2):368–377, 1984.
Brasaemle, D. L., D. M. Levin, D. C. Adler-Wailes, and C. Londos. The lipolytic stimulation of 3T3-L1 adipocytes promotes the translocation of hormone-sensitive lipase to the surfaces of lipid storage droplets. Biochim. Biophys. Acta 1483:251–262, 2000.
Bronk, B. V., G. J. Dienes, and A. Paskin. The stochastic theory of cell proliferation. Biophys. J. 8(11):1353–1398, 1968.
Chiu, Y. C., M. H. Cheng, S. Uriel, and E. M. Brey. Materials for engineering vascularized adipose tissue. J. Tissue Viabil. 20:37–48, 2011.
Conolly, R. B., and J. S. Kimbell. Computer simulation of cell growth governed by stochastic processes application to clonal growth cancer models. Toxicol. Appl. Pharmacol. 124:284–295, 1994.
Djian, P., P. Grimaldi, R. Négrel, and G. Ailhaud. Adipose conversion of Ob17 preadipocytes: relationships between cell division and fat cell cluster formation. Exp. Cell Res. 142(2):273–281, 1982.
Dominique, L. Adipose tissue lipolysis as a metabolic pathway to define pharmacological strategies against obesity and the metabolic syndrome. Pharmacol. Res. 53(6):482–491, 2006.
Forest, C., P. Grimaldi, D. Czerucka, R. Negrel, and G. Ailhaud. Establishment of a preadipocyte cell line from the epididymal fat pad of the lean C57 BL/6J mouse-long term effects of insulin and triiodothyronine on adipose conversion. In Vitro 19(4):344–354, 1983.
Geris, L., P. Van Liedekerke, B. Smeets, E. Tijskens, and H. Ramon. A cell based modelling framework for skeletal tissue engineering applications. J. Biomech. 43(5):887–892, 2010.
Green, H., and O. Kehinde. Sublines of mouse 3T3 cells that accumulate lipid. Cell 1(3):113–116, 1974.
Green, H., and O. Kehinde. An established pre-adipose cell line and its differentiation in culture II. Factors affecting the adipose conversion. Cell 5:19–25, 1975.
Green, H., and M. Meuth. An established pre-adipose cell line and its differentiation in culture. Cell 3:127–133, 1974.
Grimaldi, P., R. Negrel, J. P. Vincent, and G. Ailhaud. Differentiation of ob 17 preadipocytes to adipocytes. Effect of insulin on the levels of insulin receptors and on the transport of alpha-aminoisobutyrate. J. Biol. Chem. 254(15):6849–6852, 1979.
Gusella, J. Commitment to erythroid differentiation by friend erythroleukemia cells: a stochastic analysis. Cell 9:221–229, 1976.
Jagers, P. Stochastic models for cell kinetics. Bull. Math. Biol. 45(4):507–519, 1983.
MATLAB™. Matlab User’s Manual. Natrick, MA: The Mathworks, Inc., 2009.
Négrel, R., P. Grimaldi, and G. Ailhaud. Establishment of preadipocyte clonal line from epididymal fat pad of ob/ob mouse that responds to insulin and to lipolytic hormones. PNAS 75(12):6054–6058, 1978.
Or-Tzadikario, S., A. Gefen, et al. Confocal-based cell-specific finite element modeling extended to study variable cell shapes and intracellular structures: the example of the adipocyte. J. Biomech. 44:567–573, 2010.
Or-Tzadikario, S., R. Sopher, and A. Gefen. Quantitative monitoring of lipid accumulation over time in cultured adipocytes as function of culture conditions: toward controlled adipose tissue engineering. Tissue Eng. C 16(5):1167–1181, 2010.
Otto, T. C., and M. D. Lane. Adipose development: from stem cell to adipocyte. Crit. Rev. Biochem. Mol. Biol. 40:229–242, 2005.
Pharr, P. N., J. Nedelman, H. P. Downs, M. Ogawa, and A. J. Gross. A stochastic model for mast cell proliferation in culture. J. Cell. Physiol. 125(3):379–386, 1985.
Roeder, I., L. M. Kamminga, K. Braesel, B. Dontje, G. De Haan, and M. Loeffler. Competitive clonal hematopoiesis in mouse chimeras explained by a stochastic model of stem cell organization. Blood 105:609–616, 2005.
Shields, R., R. F. Brooks, P. N. Riddle, D. F. Capellaro, and D. Della. Cell size, cell cycle and transition probability in mouse fibroblast. Cell 15:469–474, 1978.
Shields, R., and A. Smith. Cell regulate their proliferation through alteration in transition probability. J. Cell. Physiol. 91:345–355, 1977.
Smith, J. A., and L. Martin. Do cell cycle? Proc. Natl. Acad. Sci. USA 70(4):1263–1267, 1973.
Song, H., K. C. O’connor, K. D. Papadopoulos, and D. A. Jansen. Differentiation kinetics of in vitro 3T3-L1 preadipocyte cultures. Tissue Eng. 8:1071–1081, 2002.
Steinberg, M. M., and B. L. Brownstein. Differentiation of cultured pre-adipose cells: a probability model. J. Cell Physiol. Suppl. 2:37–50, 1982.
Steppan, C. M., T. B. Shannon, B. Savitha, J. B. Elizabeth, R. B. Ronadip, M. W. Christopher, R. P. Hiralben, S. A. Rexford, and A. L. Mitchell. The hormone resistin links obesity to diabetes. Nature 409:307–312, 2000.
Vannier, C., D. Gaillard, P. Grimaldi, E. Z. Amri, P. Djian, C. Cermolacce, C. Forest, J. Etienne, R. Negrel, and G. Ailhaud. Adipose conversion of ob17 cells and hormone-related events. Int. J. Obes. 9(1):41–53, 1985.
Vogel, H., H. Niewisch, and G. Matioli. Stochastic development of stem cells. J. Theor. Biol. 22:249–270, 1969.
Yakovlev, A. Y., M. Mayer-Proschel, and M. A. Noble. A stochastic model of brain cell differentiation in tissue culture. J. Math. Biol. 37:49–60, 1998.
Acknowledgments
The authors are thankful to Ms. Shira Or-Tzadikario for the experimental data of lipid accumulation in maturing 3T3-L1 cells.
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The authors of the above paper state that they have no conflict of interest.
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Shoham, N., Gefen, A. Stochastic Modeling of Adipogenesis in 3T3-L1 Cultures to Determine Probabilities of Events in the Cell’s Life Cycle. Ann Biomed Eng 39, 2637–2653 (2011). https://doi.org/10.1007/s10439-011-0341-2
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DOI: https://doi.org/10.1007/s10439-011-0341-2