PMID- 23515914
OWN - NLM
STAT- MEDLINE
DCOM- 20140421
LR  - 20170505
IS  - 1572-8781 (Electronic)
IS  - 1387-2176 (Linking)
VI  - 15
IP  - 4
DP  - 2013 Aug
TI  - Enrichment of tumor-initiating breast cancer cells within a mammosphere-culture 
      microdevice.
PG  - 645-655
LID - 10.1007/s10544-013-9755-y [doi]
AB  - We report for the first time a microdevice that enables the selective enrichment, 
      culture, and identification of tumor-initiating cells on native 
      polydimethylsiloxane (PDMS). For nearly a decade, researchers have identified 
      tumor-initiating breast cancer cells within heterogeneous populations of breast 
      cancer cells by utilizing low-attachment serum-free culture conditions, which 
      lead to the formation of spheroidal colonies (mammospheres) that are enriched for 
      tumor-initiating cells. However, the utility of this assay has been limited by 
      difficulties in combining this culture-plate-based technique with other cellular 
      and molecular analyses. Integrating the mammosphere technique into a microsystem 
      can enable it to be combined directly with a number of functions, such as cell 
      sorting, drug screens, and molecular assays. In this work, we demonstrate 
      mammosphere culture within a PDMS microdevice. We first prove that a native 
      hydrophobic PDMS surface is as effective as commercial low-attachment plates at 
      selectively promoting the formation of mammospheres. We then experimentally 
      assess the PDMS microdevice. Time-lapse images of mammosphere formation within 
      the microdevice show that mammospheres form from single cells or small clusters 
      of cells. Following formation of the mammospheres, it is desirable to evaluate 
      the cells within the spheroids for enrichment of tumor initiating cells. To 
      perform assays such as this (which require the loading and rinsing of reagents) 
      without flushing the cells (which are in suspension) from the device, the culture 
      chamber is separated from a reagent reservoir by a commercially available 
      microporous membrane, and thus reagents are exchanged between the reservoir and 
      the culture chamber by diffusion only. Using this capability, we verify that the 
      mammospheres are enriched for tumor initiating cells by staining aldehyde 
      dehydrogenase activity, a cancer stem cell marker. To the best of our knowledge, 
      this is the first assay that enables the direct observation of tumor-initiating 
      cells within a suspended mammosphere.
FAU - Saadin, Katayoon
AU  - Saadin K
AD  - Chemical Physics Program, University of Maryland, College Park, MD, 20742, USA.
FAU - Burke, Jeffrey M
AU  - Burke JM
AD  - Fischell Department of Bioengineering, University of Maryland, College Park, MD, 
      20742, USA.
FAU - Patel, Neerav P
AU  - Patel NP
AD  - Fischell Department of Bioengineering, University of Maryland, College Park, MD, 
      20742, USA.
FAU - Zubajlo, Rebecca E
AU  - Zubajlo RE
AD  - Fischell Department of Bioengineering, University of Maryland, College Park, MD, 
      20742, USA.
FAU - White, Ian M
AU  - White IM
AD  - Chemical Physics Program, University of Maryland, College Park, MD, 20742, USA. 
      ianwhite@umd.edu.
AD  - Fischell Department of Bioengineering, University of Maryland, College Park, MD, 
      20742, USA. ianwhite@umd.edu.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biomed Microdevices
JT  - Biomedical microdevices
JID - 100887374
RN  - 0 (Biomarkers, Tumor)
RN  - 0 (Dimethylpolysiloxanes)
RN  - 0 (Indicators and Reagents)
RN  - 63148-62-9 (baysilon)
RN  - EC 1.2.1.3 (Aldehyde Dehydrogenase)
SB  - IM
MH  - Aldehyde Dehydrogenase/metabolism
MH  - Biomarkers, Tumor/metabolism
MH  - Breast Neoplasms/*pathology
MH  - Cell Culture Techniques/*instrumentation
MH  - Dimethylpolysiloxanes/chemistry
MH  - Humans
MH  - Indicators and Reagents/chemistry
MH  - MCF-7 Cells
MH  - Microtechnology/*instrumentation
MH  - Neoplastic Stem Cells/metabolism/*pathology
MH  - Reproducibility of Results
MH  - Spheroids, Cellular/*cytology
MH  - Staining and Labeling
EDAT- 2013/03/22 06:00
MHDA- 2014/04/22 06:00
CRDT- 2013/03/22 06:00
PHST- 2013/03/22 06:00 [entrez]
PHST- 2013/03/22 06:00 [pubmed]
PHST- 2014/04/22 06:00 [medline]
AID - 10.1007/s10544-013-9755-y [pii]
AID - 10.1007/s10544-013-9755-y [doi]
PST - ppublish
SO  - Biomed Microdevices. 2013 Aug;15(4):645-655. doi: 10.1007/s10544-013-9755-y.