Mass Cytometric Analysis Reveals Viable Activated Caspase-3+ Luminal Progenitors in the Normal Adult Human Mammary Gland

David J.H.F. Knapp, Nagarajan Kannan, Davide Pellacani, Connie J. Eaves

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The normal adult human female mammary gland is a bilayered structure consisting of an outer basal layer and two readily distinguished subsets of cells within the inner luminal layer. We now present a validated methodology for undertaking large-scale multi-parameter mass cytometric analyses of these cell types at single-cell resolution. In addition, we show how combining this approach with in vitro clonogenic assays of the proliferative and signaling responses of normal human mammary cells to epidermal growth factor (EGF) allows additional subsets with different EGF responses to be discerned. This included the identification of a subset of cells within the phenotypically defined luminal progenitor fraction that displays an elevated content of active caspase-3, including some that generate clones in vitro in response to EGF, with immunohistochemical evidence of their presence in situ in fixed preparations of normal human breast tissue. Knapp et al. identify a subpopulation of progenitors in the human mammary gland that have partially activated an apoptotic program despite retaining some viability and clonogenic potential. As such activation has been linked to genomic instability, these cells may be at increased risk for oncogenic transformation.

Original languageEnglish (US)
Pages (from-to)1116-1126
Number of pages11
JournalCell Reports
Volume21
Issue number4
DOIs
StatePublished - Oct 24 2017

Fingerprint

Human Mammary Glands
Epidermal Growth Factor
Caspase 3
Breast
Assays
Single-Cell Analysis
Chemical activation
Tissue
Genomic Instability
Clone Cells
In Vitro Techniques

Keywords

  • apoptosis
  • cancer
  • cell death
  • genomic instability
  • growth factors
  • mammary
  • mass cytometry
  • signaling
  • single-cell biology

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Mass Cytometric Analysis Reveals Viable Activated Caspase-3+ Luminal Progenitors in the Normal Adult Human Mammary Gland. / Knapp, David J.H.F.; Kannan, Nagarajan; Pellacani, Davide; Eaves, Connie J.

In: Cell Reports, Vol. 21, No. 4, 24.10.2017, p. 1116-1126.

Research output: Contribution to journalArticle

Knapp, David J.H.F. ; Kannan, Nagarajan ; Pellacani, Davide ; Eaves, Connie J. / Mass Cytometric Analysis Reveals Viable Activated Caspase-3+ Luminal Progenitors in the Normal Adult Human Mammary Gland. In: Cell Reports. 2017 ; Vol. 21, No. 4. pp. 1116-1126.
@article{2310dd3d4ff244f196b4e48f4dd8c6fd,
title = "Mass Cytometric Analysis Reveals Viable Activated Caspase-3+ Luminal Progenitors in the Normal Adult Human Mammary Gland",
abstract = "The normal adult human female mammary gland is a bilayered structure consisting of an outer basal layer and two readily distinguished subsets of cells within the inner luminal layer. We now present a validated methodology for undertaking large-scale multi-parameter mass cytometric analyses of these cell types at single-cell resolution. In addition, we show how combining this approach with in vitro clonogenic assays of the proliferative and signaling responses of normal human mammary cells to epidermal growth factor (EGF) allows additional subsets with different EGF responses to be discerned. This included the identification of a subset of cells within the phenotypically defined luminal progenitor fraction that displays an elevated content of active caspase-3, including some that generate clones in vitro in response to EGF, with immunohistochemical evidence of their presence in situ in fixed preparations of normal human breast tissue. Knapp et al. identify a subpopulation of progenitors in the human mammary gland that have partially activated an apoptotic program despite retaining some viability and clonogenic potential. As such activation has been linked to genomic instability, these cells may be at increased risk for oncogenic transformation.",
keywords = "apoptosis, cancer, cell death, genomic instability, growth factors, mammary, mass cytometry, signaling, single-cell biology",
author = "Knapp, {David J.H.F.} and Nagarajan Kannan and Davide Pellacani and Eaves, {Connie J.}",
year = "2017",
month = "10",
day = "24",
doi = "10.1016/j.celrep.2017.09.096",
language = "English (US)",
volume = "21",
pages = "1116--1126",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "4",

}

TY - JOUR

T1 - Mass Cytometric Analysis Reveals Viable Activated Caspase-3+ Luminal Progenitors in the Normal Adult Human Mammary Gland

AU - Knapp, David J.H.F.

AU - Kannan, Nagarajan

AU - Pellacani, Davide

AU - Eaves, Connie J.

PY - 2017/10/24

Y1 - 2017/10/24

N2 - The normal adult human female mammary gland is a bilayered structure consisting of an outer basal layer and two readily distinguished subsets of cells within the inner luminal layer. We now present a validated methodology for undertaking large-scale multi-parameter mass cytometric analyses of these cell types at single-cell resolution. In addition, we show how combining this approach with in vitro clonogenic assays of the proliferative and signaling responses of normal human mammary cells to epidermal growth factor (EGF) allows additional subsets with different EGF responses to be discerned. This included the identification of a subset of cells within the phenotypically defined luminal progenitor fraction that displays an elevated content of active caspase-3, including some that generate clones in vitro in response to EGF, with immunohistochemical evidence of their presence in situ in fixed preparations of normal human breast tissue. Knapp et al. identify a subpopulation of progenitors in the human mammary gland that have partially activated an apoptotic program despite retaining some viability and clonogenic potential. As such activation has been linked to genomic instability, these cells may be at increased risk for oncogenic transformation.

AB - The normal adult human female mammary gland is a bilayered structure consisting of an outer basal layer and two readily distinguished subsets of cells within the inner luminal layer. We now present a validated methodology for undertaking large-scale multi-parameter mass cytometric analyses of these cell types at single-cell resolution. In addition, we show how combining this approach with in vitro clonogenic assays of the proliferative and signaling responses of normal human mammary cells to epidermal growth factor (EGF) allows additional subsets with different EGF responses to be discerned. This included the identification of a subset of cells within the phenotypically defined luminal progenitor fraction that displays an elevated content of active caspase-3, including some that generate clones in vitro in response to EGF, with immunohistochemical evidence of their presence in situ in fixed preparations of normal human breast tissue. Knapp et al. identify a subpopulation of progenitors in the human mammary gland that have partially activated an apoptotic program despite retaining some viability and clonogenic potential. As such activation has been linked to genomic instability, these cells may be at increased risk for oncogenic transformation.

KW - apoptosis

KW - cancer

KW - cell death

KW - genomic instability

KW - growth factors

KW - mammary

KW - mass cytometry

KW - signaling

KW - single-cell biology

UR - http://www.scopus.com/inward/record.url?scp=85032026656&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85032026656&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2017.09.096

DO - 10.1016/j.celrep.2017.09.096

M3 - Article

C2 - 29069592

AN - SCOPUS:85032026656

VL - 21

SP - 1116

EP - 1126

JO - Cell Reports

JF - Cell Reports

SN - 2211-1247

IS - 4

ER -