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Integr Biol (Camb). 2014 Feb;6(2):224-31. doi: 10.1039/c3ib40136g.

Streamlining gene expression analysis: integration of co-culture and mRNA purification.

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1
Departments of Biomedical Engineering, University of Wisconsin-Madison, 1111 Highland Ave., Madison, WI 53705, USA. Berry3@wisc.edu.

Abstract

Co-culture of multiple cell types within a single device enables the study of paracrine signaling events. However, extracting gene expression endpoints from co-culture experiments is laborious, due in part to pre-PCR processing of the sample (i.e., post-culture cell sorting and nucleic acid purification). Also, a significant loss of nucleic acid may occur during these steps, especially with microfluidic cell culture where lysate volumes are small and difficult to access. Here, we describe an integrated platform for performing microfluidic cell culture and extraction of mRNA for gene expression analysis. This platform was able to recover 30-fold more mRNA than a similar, non-integrated system. Additionally, using a breast cancer/bone marrow stroma co-culture, we recapitulated stromal-dependent, estrogen-independent growth of the breast cancer cells, coincident with transcriptional changes. We anticipate that this platform will be used for streamlined analysis of paracrine signaling events as well as for screening potential drugs and/or patient samples.

PMID:
24413730
PMCID:
PMC3956049
DOI:
10.1039/c3ib40136g
[Indexed for MEDLINE]
Free PMC Article
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