Quantitative analysis by flow cytometry of interstitial cells of Cajal, pacemakers, and mediators of neurotransmission in the gastrointestinal tract

Tamas Ordog, Doug Redelman, Viktor J. Horváth, Lisa J. Miller, Burton Horowitz, Kenton M. Sanders

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Background: Interstitial cells of Cajal (ICCs) are mesenchymal cells that play critical roles in gastrointestinal motility as electrical pacemakers and mediators of neuromuscular neurotransmission. Although depletions of ICCs have been implicated in several gastrointestinal motor disorders, quantification of these cells has been difficult due to their varied morphology, regionally changing network density, and overall scarcity. Our goal was to evaluate flow cytometry (FCM) for the enumeration of ICCs. Methods: We identified murine ICCs in live gastrointestinal muscles or primary cell cultures grown in the presence or absence of stem cell factor (SCF)- expressing STO fibroblasts with fluorescent Kit (CD117) antibodies. Because this technique also labels resident macrophages nonspecifically, we identified the latter with additional fluorescent antibodies. Dispersed cells were analyzed by FCM. Results: ICCs represented 1.63 ± 0.17% of the total cell count in the distal stomach (n = 18 mice) and 5.85 ± 0.84% in the proximal colon and 6.28 ± 0.61% in the distal colon (n = 3 mice). In fundic muscles of W/Wv mice (n = 5) that virtually lack ICCs, very few Kit+ cells were detected. FCM identified approximately 2.6- to 7.3-fold more Kit+ ICCs in small intestinal cell cultures grown on STO flbroblasts expressing membrane-bound SCF (n = 6) than in cultures stimulated with soluble SCF (n = 6). Conclusions: FCM is a sensitive and specific method for the unbiased quantification of ICCs.

Original languageEnglish (US)
Pages (from-to)139-149
Number of pages11
JournalCytometry Part A
Volume62
Issue number2
DOIs
StatePublished - Dec 2004
Externally publishedYes

Fingerprint

Interstitial Cells of Cajal
Synaptic Transmission
Gastrointestinal Tract
Flow Cytometry
Stem Cell Factor
Colon
Muscles
Gastrointestinal Motility
Primary Cell Culture
Antibodies
Stomach
Cell Culture Techniques
Fibroblasts
Cell Count
Macrophages
Membranes

Keywords

  • CD117
  • Flow cytometry
  • Immunofluorescence
  • Interstitial cells of Cajal
  • Kit
  • Macrophage
  • Mouse
  • Quantitative reverse transcription-polymerase chain reaction
  • Stem cell factor
  • STO fibroblast

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Quantitative analysis by flow cytometry of interstitial cells of Cajal, pacemakers, and mediators of neurotransmission in the gastrointestinal tract. / Ordog, Tamas; Redelman, Doug; Horváth, Viktor J.; Miller, Lisa J.; Horowitz, Burton; Sanders, Kenton M.

In: Cytometry Part A, Vol. 62, No. 2, 12.2004, p. 139-149.

Research output: Contribution to journalArticle

Ordog, Tamas ; Redelman, Doug ; Horváth, Viktor J. ; Miller, Lisa J. ; Horowitz, Burton ; Sanders, Kenton M. / Quantitative analysis by flow cytometry of interstitial cells of Cajal, pacemakers, and mediators of neurotransmission in the gastrointestinal tract. In: Cytometry Part A. 2004 ; Vol. 62, No. 2. pp. 139-149.
@article{6324602e027f46d5af19c6893be02f9d,
title = "Quantitative analysis by flow cytometry of interstitial cells of Cajal, pacemakers, and mediators of neurotransmission in the gastrointestinal tract",
abstract = "Background: Interstitial cells of Cajal (ICCs) are mesenchymal cells that play critical roles in gastrointestinal motility as electrical pacemakers and mediators of neuromuscular neurotransmission. Although depletions of ICCs have been implicated in several gastrointestinal motor disorders, quantification of these cells has been difficult due to their varied morphology, regionally changing network density, and overall scarcity. Our goal was to evaluate flow cytometry (FCM) for the enumeration of ICCs. Methods: We identified murine ICCs in live gastrointestinal muscles or primary cell cultures grown in the presence or absence of stem cell factor (SCF)- expressing STO fibroblasts with fluorescent Kit (CD117) antibodies. Because this technique also labels resident macrophages nonspecifically, we identified the latter with additional fluorescent antibodies. Dispersed cells were analyzed by FCM. Results: ICCs represented 1.63 ± 0.17{\%} of the total cell count in the distal stomach (n = 18 mice) and 5.85 ± 0.84{\%} in the proximal colon and 6.28 ± 0.61{\%} in the distal colon (n = 3 mice). In fundic muscles of W/Wv mice (n = 5) that virtually lack ICCs, very few Kit+ cells were detected. FCM identified approximately 2.6- to 7.3-fold more Kit+ ICCs in small intestinal cell cultures grown on STO flbroblasts expressing membrane-bound SCF (n = 6) than in cultures stimulated with soluble SCF (n = 6). Conclusions: FCM is a sensitive and specific method for the unbiased quantification of ICCs.",
keywords = "CD117, Flow cytometry, Immunofluorescence, Interstitial cells of Cajal, Kit, Macrophage, Mouse, Quantitative reverse transcription-polymerase chain reaction, Stem cell factor, STO fibroblast",
author = "Tamas Ordog and Doug Redelman and Horv{\'a}th, {Viktor J.} and Miller, {Lisa J.} and Burton Horowitz and Sanders, {Kenton M.}",
year = "2004",
month = "12",
doi = "10.1002/cyto.a.20078",
language = "English (US)",
volume = "62",
pages = "139--149",
journal = "Cytometry. Part A : the journal of the International Society for Analytical Cytology",
issn = "1552-4922",
publisher = "Wiley-Liss Inc.",
number = "2",

}

TY - JOUR

T1 - Quantitative analysis by flow cytometry of interstitial cells of Cajal, pacemakers, and mediators of neurotransmission in the gastrointestinal tract

AU - Ordog, Tamas

AU - Redelman, Doug

AU - Horváth, Viktor J.

AU - Miller, Lisa J.

AU - Horowitz, Burton

AU - Sanders, Kenton M.

PY - 2004/12

Y1 - 2004/12

N2 - Background: Interstitial cells of Cajal (ICCs) are mesenchymal cells that play critical roles in gastrointestinal motility as electrical pacemakers and mediators of neuromuscular neurotransmission. Although depletions of ICCs have been implicated in several gastrointestinal motor disorders, quantification of these cells has been difficult due to their varied morphology, regionally changing network density, and overall scarcity. Our goal was to evaluate flow cytometry (FCM) for the enumeration of ICCs. Methods: We identified murine ICCs in live gastrointestinal muscles or primary cell cultures grown in the presence or absence of stem cell factor (SCF)- expressing STO fibroblasts with fluorescent Kit (CD117) antibodies. Because this technique also labels resident macrophages nonspecifically, we identified the latter with additional fluorescent antibodies. Dispersed cells were analyzed by FCM. Results: ICCs represented 1.63 ± 0.17% of the total cell count in the distal stomach (n = 18 mice) and 5.85 ± 0.84% in the proximal colon and 6.28 ± 0.61% in the distal colon (n = 3 mice). In fundic muscles of W/Wv mice (n = 5) that virtually lack ICCs, very few Kit+ cells were detected. FCM identified approximately 2.6- to 7.3-fold more Kit+ ICCs in small intestinal cell cultures grown on STO flbroblasts expressing membrane-bound SCF (n = 6) than in cultures stimulated with soluble SCF (n = 6). Conclusions: FCM is a sensitive and specific method for the unbiased quantification of ICCs.

AB - Background: Interstitial cells of Cajal (ICCs) are mesenchymal cells that play critical roles in gastrointestinal motility as electrical pacemakers and mediators of neuromuscular neurotransmission. Although depletions of ICCs have been implicated in several gastrointestinal motor disorders, quantification of these cells has been difficult due to their varied morphology, regionally changing network density, and overall scarcity. Our goal was to evaluate flow cytometry (FCM) for the enumeration of ICCs. Methods: We identified murine ICCs in live gastrointestinal muscles or primary cell cultures grown in the presence or absence of stem cell factor (SCF)- expressing STO fibroblasts with fluorescent Kit (CD117) antibodies. Because this technique also labels resident macrophages nonspecifically, we identified the latter with additional fluorescent antibodies. Dispersed cells were analyzed by FCM. Results: ICCs represented 1.63 ± 0.17% of the total cell count in the distal stomach (n = 18 mice) and 5.85 ± 0.84% in the proximal colon and 6.28 ± 0.61% in the distal colon (n = 3 mice). In fundic muscles of W/Wv mice (n = 5) that virtually lack ICCs, very few Kit+ cells were detected. FCM identified approximately 2.6- to 7.3-fold more Kit+ ICCs in small intestinal cell cultures grown on STO flbroblasts expressing membrane-bound SCF (n = 6) than in cultures stimulated with soluble SCF (n = 6). Conclusions: FCM is a sensitive and specific method for the unbiased quantification of ICCs.

KW - CD117

KW - Flow cytometry

KW - Immunofluorescence

KW - Interstitial cells of Cajal

KW - Kit

KW - Macrophage

KW - Mouse

KW - Quantitative reverse transcription-polymerase chain reaction

KW - Stem cell factor

KW - STO fibroblast

UR - http://www.scopus.com/inward/record.url?scp=9644268794&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=9644268794&partnerID=8YFLogxK

U2 - 10.1002/cyto.a.20078

DO - 10.1002/cyto.a.20078

M3 - Article

C2 - 15536638

AN - SCOPUS:9644268794

VL - 62

SP - 139

EP - 149

JO - Cytometry. Part A : the journal of the International Society for Analytical Cytology

JF - Cytometry. Part A : the journal of the International Society for Analytical Cytology

SN - 1552-4922

IS - 2

ER -