MDCK Cystogenesis Driven by Cell Stabilization within Computational Analogues

University of California San Diego, United States of America
PLoS Computational Biology (Impact Factor: 4.83). 04/2011; 7(4):e1002030. DOI: 10.1371/journal.pcbi.1002030
Source: PubMed

ABSTRACT The study of epithelial morphogenesis is fundamental to increasing our understanding of organ function and disease. Great progress has been made through study of culture systems such as Madin-Darby canine kidney (MDCK) cells, but many aspects of even simple morphogenesis remain unclear. For example, are specific cell actions tightly coupled to the characteristics of the cell's environment or are they more often cell state dependent? How does the single lumen, single cell layer cyst consistently emerge from a variety of cell actions? To improve insight, we instantiated in silico analogues that used hypothesized cell behavior mechanisms to mimic MDCK cystogenesis. We tested them through in vitro experimentation and quantitative validation. We observed novel growth patterns, including a cell behavior shift that began around day five of growth. We created agent-oriented analogues that used the cellular Potts model along with an Iterative Refinement protocol. Following several refinements, we achieved a degree of validation for two separate mechanisms. Both survived falsification and achieved prespecified measures of similarity to cell culture properties. In silico components and mechanisms mapped to in vitro counterparts. In silico, the axis of cell division significantly affects lumen number without changing cell number or cyst size. Reducing the amount of in silico luminal cell death had limited effect on cystogenesis. Simulations provide an observable theory for cystogenesis based on hypothesized, cell-level operating principles.

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Available from: Jesse A Engelberg, Aug 12, 2015
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    • "In particular , it has not been demonstrated whether the simple aforementioned mechanistic principles are sufficient to reproduce cyst architectures. Furthermore, although it is clear that misorientation of the spindle is an ingredient leading to the aberrant multilumen phenotype (Zheng et al., 2010; Engelberg et al., 2011), it is not clear how and to what extent cyst architecture and orientation of cell division are related. To shed light on these issues, we developed a mathematical model based on the phenomenology of cell–cell and cell– matrix interactions that reproduces the experimentally observed phenotypes in both the normal and the aberrant cases. "
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    ABSTRACT: The growth of a well-formed epithelial structure is governed by mechanical constraints, cellular apico-basal polarity, and spatially controlled cell division. Here we compared the predictions of a mathematical model of epithelial growth with the morphological analysis of 3D epithelial structures. In both in vitro cyst models and in developing epithelial structures in vivo, epithelial growth could take place close to or far from mechanical equilibrium, and was determined by the hierarchy of time-scales of cell division, cell-cell rearrangements, and lumen dynamics. Equilibrium properties could be inferred by the analysis of cell-cell contact topologies, and the nonequilibrium phenotype was altered by inhibiting ROCK activity. The occurrence of an aberrant multilumen phenotype was linked to fast nonequilibrium growth, even when geometric control of cell division was correctly enforced. We predicted and verified experimentally that slowing down cell division partially rescued a multilumen phenotype induced by altered polarity. These results improve our understanding of the development of epithelial organs and, ultimately, of carcinogenesis.
    The Journal of Cell Biology 10/2013; 203(2). DOI:10.1083/jcb.201305044 · 9.69 Impact Factor
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    • "The molecular mechanisms of microlumen formation have been intensively studied using in vitro cell culture systems, such as the MDCK (Madin-Darby canine kidney) cell line (Wang et al. 1990; Apodaca 2010; Engelberg et al. 2011). By culturing three-dimensionally in a collagen gel, individual MDCK cells proliferate and assemble into a cyst, which is a spherical epithelial layer surrounding a central cavity. "
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    ABSTRACT: Trophectoderm (TE) is the first cell type that emerges during development and plays pivotal roles in the viviparous mode of reproduction in placental mammals. TE adopts typical epithelium morphology to surround a fluid-filled cavity, whose expansion is critical for hatching and efficient interaction with the uterine endometrium for implantation. TE also differentiates into trophoblast cells to construct the placenta. This chapter is an overview of the cellular and molecular mechanisms that control the critical aspects of TE formation, namely, the formation of the blastocyst cavity, the expression of key transcription factors, and the roles of cell polarity in the specification of the TE lineage. Current gaps in our knowledge and challenging issues are also discussed that should be addressed in future investigations in order to further advance our understanding of the mechanisms of TE formation.
    Results and problems in cell differentiation 01/2012; 55:165-84. DOI:10.1007/978-3-642-30406-4_9
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    • "For example, CELL DIVI- SION was implemented in an agent-based manner so CELLS had access to information about the points contained within them, thus it was straightforward to randomize or invert the axis of CELL DIVISION. In order to validate the quantitative results of ISMA-M and ISMA-C, similarity measures were developed for cell number, lumen and cyst size, mean cell area, and the ratio of cellular to cyst area [2]. Similarity Measure 1 (SM1) measured the percentage of in silico simulations at a given day that were within ± 25% of the mean in vitro value at that day, "
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    ABSTRACT: Madin-Darby canine kidney (MDCK) cells undergoing cystogenesis in vitro is a scientifically useful model of epithelial morphogenesis. The cysts formed in collagen and Matrigel are qualitatively similar, consisting of a single layer of epithelial cells surrounding a hollow lumen. However, differences in key quantitative measures of cyst growth, including cell number and cyst and lumen size, indicate that some cell behaviors are different within the two culture systems. We recently described an agent-oriented, agent-directed analogue of MDCK cystogenesis in Matrigel. It utilized a cellular Potts model and achieved qualitative and quantitative validation targets using empirical parameter tuning. Within this report we highlight steps taken to convert the cellular Potts model framework to one based upon an agent-oriented approach. If measures of cell death are ignored, the only parameters that required adjustment to allow the analogue of cystogenesis in Matrigel to mimic MDCK cystogenesis in collagen were those controlling cell division and polarization. These data indicate that in addition to delayed cell polarization, cell division in collagen is likely slower than in Matrigel. The reported results strongly support the hypothesis that MDCK cells use the same basic operating principles to create cysts when cultured in Matrigel or collagen.
    2011 Spring Simulation Multi-conference, SpringSim '11, Boston, MA, USA, April 03-07, 2011. Volume 1: Proceedings of the 2011 Workshop on Agent-Directed Simulation (ADS).; 01/2011
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