title: Transitions from mono- to co- to tri-culture uniquely affect gene expression in breast cancer, stromal, and immune compartments.
abstract: Heterotypic interactions in cancer microenvironments play important roles in disease initiation, progression, and spread. Co-culture is the predominant approach used in dissecting paracrine interactions between tumor and stromal cells, but functional results from simple co-cultures frequently fail to correlate to in vivo conditions. Though complex heterotypic in vitro models have improved functional relevance, there is little systematic knowledge of how multi-culture parameters influence this recapitulation. We therefore have employed a more iterative approach to investigate the influence of increasing model complexity; increased heterotypic complexity specifically. Here we describe how the compartmentalized and microscale elements of our multi-culture device allowed us to obtain gene expression data from one cell type at a time in a heterotypic culture where cells communicated through paracrine interactions. With our device we generated a large dataset comprised of cell type specific gene-expression patterns for cultures of increasing complexity (three cell types in mono-, co-, or tri-culture) not readily accessible in other systems. Principal component analysis indicated that gene expression was changed in co-culture but was often more strongly altered in tri-culture as compared to mono-culture. Our analysis revealed that cell type identity and the complexity around it (mono-, co-, or tri-culture) influence gene regulation. We also observed evidence of complementary regulation between cell types in the same heterotypic culture. Here we demonstrate the utility of our platform in providing insight into how tumor and stromal cells respond to microenvironments of varying complexities highlighting the expanding importance of heterotypic cultures that go beyond conventional co-culture.
authors.affiliation: Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Mary C
authors.initials: MC
authors.lastname: Regier
authors.affiliation: Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.|Department of Oncology, McArdle Laboratories for Cancer Research, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Lindsey J
authors.initials: LJ
authors.lastname: Maccoux
authors.affiliation: Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Emma M
authors.initials: EM
authors.lastname: Weinberger
authors.affiliation: Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Keil J
authors.initials: KJ
authors.lastname: Regehr
authors.affiliation: Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Berry
authors.affiliation: Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.|University of Wisconsin Carbone Cancer Center, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
authors.affiliation: Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.|Department of Oncology, McArdle Laboratories for Cancer Research, University of Wisconsin-Madison, Madison, WI, USA.|University of Wisconsin Carbone Cancer Center, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Elaine T
authors.initials: ET
authors.lastname: Alarid
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics*; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D025341:Principal Component Analysis; D017154:Stromal Cells / Q000378:metabolism*; D059016:Tumor Microenvironment
keywords: Compartmentalization; Heterotypic interactions; Microfluidic; Multi-culture; Principal component analysis