Longitudinal Bioluminescent Quantification of Three Dimensional Cell Growth

Michael K Wendt; William P Schiemann

doi:10.21769/BioProtoc.993

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Longitudinal Bioluminescent Quantification of Three Dimensional Cell Growth

Journal article

Open access

Peer reviewed

Longitudinal Bioluminescent Quantification of Three Dimensional Cell Growth

Michael K Wendt and William P Schiemann

Bio-protocol, Vol.3(23), e993

12/05/2013

DOI: 10.21769/BioProtoc.993

PMCID: PMC4957652

PMID: 27453906

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https://doi.org/10.21769/BioProtoc.993View

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Abstract

The use of three-dimensional (3D) cell culture systems is widely accepted as representing a more physiologically relevant means to propagate mammary epithelial and breast cancer cells. However, 3D cultures systems are plagued by several experimental and technical limitations as compared to their traditional 2D counterparts. For instance, quantifying the growth of mammary epithelial or breast cancer organoids longitudinally is particularly troublesome using standard [ H]thymidine or MTT assay systems, or using computer-assisted area calculations. Likewise, the nature of the multicellular aggregates and organoids formed by breast cancer cells under 3D conditions precludes efficient recovery of the cells from 3D matrices, an event that is time consuming and leads to spurious results. The assay described here utilizes stable expression of firefly luciferase as means to quantify the longitudinal outgrowth of cells propagated within a 3D matrices. The major advantages of this technique include its high-throughput nature and ability to longitudinally track single wells over a defined period of time, thereby decreasing the costs associated with assay performance. Finally, this technique can be readily combined with drug treatments and/or genetic manipulations to assay their effects on the growth of 3D organoids.

Details

Title: Subtitle: Longitudinal Bioluminescent Quantification of Three Dimensional Cell Growth
Creators: Michael K Wendt - Case Western Reserve University
William P Schiemann - Case Western Reserve University
Resource Type: Journal article
Publication Details: Bio-protocol, Vol.3(23), e993
DOI: 10.21769/BioProtoc.993
PMID: 27453906
PMCID: PMC4957652
ISSN: 2331-8325
eISSN: 2331-8325
Grant note: R01 CA177069 / NCI NIH HHS R00 CA166140 / NCI NIH HHS R01 CA129359 / NCI NIH HHS K99 CA166140 / NCI NIH HHS
Language: English
Date published: 12/05/2013
Academic Unit: Internal Medicine
Record Identifier: 9984479264302771

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