PMID- 23567076
OWN - NLM
STAT- MEDLINE
DCOM- 20140210
LR  - 20130710
IS  - 1873-488X (Electronic)
IS  - 1056-8719 (Linking)
VI  - 68
IP  - 1
DP  - 2013 Jul-Aug
TI  - Minimized cell usage for stem cell-derived and primary cells on an automated 
      patch clamp system.
PG  - 82-7
LID - S1056-8719(13)00232-3 [pii]
LID - 10.1016/j.vascn.2013.03.009 [doi]
AB  - INTRODUCTION: Chip-based automated patch clamp systems are widely used in drug 
      development and safety pharmacology, allowing for high quality, high throughput 
      screening at standardized experimental conditions. The merits of automation 
      generally come at the cost of large amounts of cells needed, since cells are not 
      targeted individually, but randomly positioned onto the chip aperture from cells 
      in suspension. While cell usage is of little concern when using standard cell 
      lines such as CHO or HEK cells, it becomes a crucial constraint with cells of 
      limited availability, such as primary or otherwise rare and expensive cells, like 
      induced pluripotent stem (IPS) cell-derived cardiomyocytes or neurons. METHODS: 
      We established application protocols for CHO cells, IPS cell-derived neurons 
      (iCell(R) Neurons, Cellular Dynamics International), cardiomyocytes (Cor.4U(R), 
      Axiogenesis) and pancreatic islet cells, minimizing cell usage for automated 
      patch clamp recordings on Nanion's Patchliner. Use of 5 mul cell suspension per 
      well for densities between 55,000 cells/ml and 400,000 cells/ml depending on cell 
      type resulted in good cell capture. RESULTS: We present a new cell application 
      procedure optimized for the Patchliner achieving>80% success rates for using as 
      little as 300 to 2000 cells per well depending on cell type. We demonstrate that 
      this protocol works for standard cell lines, as well as for stem cell-derived 
      neurons and cardiomyocytes, and for primary pancreatic islet cells. We present 
      recordings for these cell types, demonstrating that high data quality is not 
      compromised by altered cell application. DISCUSSION: Our new cell application 
      procedure achieves high success rates with unprecedentedly low cell numbers. 
      Compared to other standard automated patch clamp systems we reduced the average 
      amount of cells needed by more than 150 times. Reduced cell usage crucially 
      improves cost efficiency for expensive cells and opens up automated patch clamp 
      for primary cells of limited availability.
CI  - Copyright (c) 2013 Elsevier Inc. All rights reserved.
FAU - Becker, Nadine
AU  - Becker N
AD  - Nanion Technologies GmbH, Gabrielenstrasse 9, 80636 Munich, Germany. 
      nadine@nanion.de
FAU - Stoelzle, Sonja
AU  - Stoelzle S
FAU - Gopel, Sven
AU  - Gopel S
FAU - Guinot, David
AU  - Guinot D
FAU - Mumm, Patrick
AU  - Mumm P
FAU - Haarmann, Claudia
AU  - Haarmann C
FAU - Malan, Daniela
AU  - Malan D
FAU - Bohlen, Heribert
AU  - Bohlen H
FAU - Kossolov, Eugen
AU  - Kossolov E
FAU - Kettenhofen, Ralf
AU  - Kettenhofen R
FAU - George, Michael
AU  - George M
FAU - Fertig, Niels
AU  - Fertig N
FAU - Bruggemann, Andrea
AU  - Bruggemann A
LA  - eng
PT  - Comparative Study
PT  - Journal Article
DEP - 20130406
PL  - United States
TA  - J Pharmacol Toxicol Methods
JT  - Journal of pharmacological and toxicological methods
JID - 9206091
SB  - IM
MH  - Animals
MH  - Automation
MH  - CHO Cells/cytology
MH  - Cricetinae
MH  - Cricetulus
MH  - Humans
MH  - Induced Pluripotent Stem Cells/*cytology
MH  - Islets of Langerhans/cytology
MH  - Mice
MH  - Myocytes, Cardiac/*cytology
MH  - Neurons/*cytology
MH  - Patch-Clamp Techniques/economics/*methods
OTO - NOTNLM
OT  - Automated patch clamp
OT  - Patchliner
OT  - pancreatic islet cells
OT  - primary cells
OT  - stem cell-derived cardiomyocytes
OT  - stem cell-derived neurons
EDAT- 2013/04/10 06:00
MHDA- 2014/02/11 06:00
CRDT- 2013/04/10 06:00
PHST- 2013/02/08 00:00 [received]
PHST- 2013/03/27 00:00 [revised]
PHST- 2013/03/27 00:00 [accepted]
PHST- 2013/04/10 06:00 [entrez]
PHST- 2013/04/10 06:00 [pubmed]
PHST- 2014/02/11 06:00 [medline]
AID - S1056-8719(13)00232-3 [pii]
AID - 10.1016/j.vascn.2013.03.009 [doi]
PST - ppublish
SO  - J Pharmacol Toxicol Methods. 2013 Jul-Aug;68(1):82-7. doi: 
      10.1016/j.vascn.2013.03.009. Epub 2013 Apr 6.