Culture requirements of prostatic epithelial cell lines for acinar morphogenesis and lumen formation in vitro: Role of extracellular calcium

Darren R. Tyson, Junichi Inokuchi, Toshiyuki Tsunoda, Alice Lau, David K. Ornstein

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

BACKGROUND. Three-dimensional (3D) culture of benign prostatic epithelial cell lines can recapitulate acinar morphogenesis in vitro, but the broad applicability of this approach has not been described. The present studies examine the culture conditions important for prostatic acinar morphogenesis in vitro and the role of extracellular calcium in this process. METHODS. With optimized culture conditions, RWPE-1, pRNS-1-1, PZ-HPV-7, PNT1A, BPH-1, and PrEC were analyzed for their ability to undergo acinar morphogenesis in 3D culture and by immunoblotting. RWPE-1 cells were further examined for the effects of calcium on morphology, E-cadherin membrane localization and multicellular layering in 2D culture and for acinar morphogenesis, luminal apoptosis, and luminal filling in 3D. RESULTS. Cell lines grown in low-calcium medium have the ability to form acinar structures with lumens, which correlates with E-cadherin expression, but low calcium is not required for this process. Adding CaCl 2 to the medium strongly inhibits lumen formation, luminal apoptosis and induces luminal filling, and luminal filling is blocked by an interfering antibody. CONCLUSIONS. Optimized medium composition allows nearly all seeded RWPE-1 cells to undergo acinar morphogenesis, forming consistent structures representative of normal adult prostate glands. Low-calcium-containing medium appears selective for cells capable of undergoing acinar morphogenesis in vitro, and branching and luminal space within the acini are strongly influenced by extracellular calcium levels, likely through the actions of E-cadherin. These results provide important information about a relevant in vitro model with which to study prostate development and carcinogenesis and highlight the importance of extracellular calcium in regulating 3D morphology.

Original languageEnglish
Pages (from-to)1601-1613
Number of pages13
JournalProstate
Volume67
Issue number15
DOIs
Publication statusPublished - Nov 1 2007

Fingerprint

Morphogenesis
Epithelial Cells
Calcium
Cell Line
Cadherins
Prostate
Apoptosis
In Vitro Techniques
Immunoblotting
Carcinogenesis
Membranes
Antibodies

All Science Journal Classification (ASJC) codes

  • Oncology
  • Urology

Cite this

Culture requirements of prostatic epithelial cell lines for acinar morphogenesis and lumen formation in vitro : Role of extracellular calcium. / Tyson, Darren R.; Inokuchi, Junichi; Tsunoda, Toshiyuki; Lau, Alice; Ornstein, David K.

In: Prostate, Vol. 67, No. 15, 01.11.2007, p. 1601-1613.

Research output: Contribution to journalArticle

Tyson, Darren R. ; Inokuchi, Junichi ; Tsunoda, Toshiyuki ; Lau, Alice ; Ornstein, David K. / Culture requirements of prostatic epithelial cell lines for acinar morphogenesis and lumen formation in vitro : Role of extracellular calcium. In: Prostate. 2007 ; Vol. 67, No. 15. pp. 1601-1613.
@article{c393a7531359427c928a36f174b3bc2c,
title = "Culture requirements of prostatic epithelial cell lines for acinar morphogenesis and lumen formation in vitro: Role of extracellular calcium",
abstract = "BACKGROUND. Three-dimensional (3D) culture of benign prostatic epithelial cell lines can recapitulate acinar morphogenesis in vitro, but the broad applicability of this approach has not been described. The present studies examine the culture conditions important for prostatic acinar morphogenesis in vitro and the role of extracellular calcium in this process. METHODS. With optimized culture conditions, RWPE-1, pRNS-1-1, PZ-HPV-7, PNT1A, BPH-1, and PrEC were analyzed for their ability to undergo acinar morphogenesis in 3D culture and by immunoblotting. RWPE-1 cells were further examined for the effects of calcium on morphology, E-cadherin membrane localization and multicellular layering in 2D culture and for acinar morphogenesis, luminal apoptosis, and luminal filling in 3D. RESULTS. Cell lines grown in low-calcium medium have the ability to form acinar structures with lumens, which correlates with E-cadherin expression, but low calcium is not required for this process. Adding CaCl 2 to the medium strongly inhibits lumen formation, luminal apoptosis and induces luminal filling, and luminal filling is blocked by an interfering antibody. CONCLUSIONS. Optimized medium composition allows nearly all seeded RWPE-1 cells to undergo acinar morphogenesis, forming consistent structures representative of normal adult prostate glands. Low-calcium-containing medium appears selective for cells capable of undergoing acinar morphogenesis in vitro, and branching and luminal space within the acini are strongly influenced by extracellular calcium levels, likely through the actions of E-cadherin. These results provide important information about a relevant in vitro model with which to study prostate development and carcinogenesis and highlight the importance of extracellular calcium in regulating 3D morphology.",
author = "Tyson, {Darren R.} and Junichi Inokuchi and Toshiyuki Tsunoda and Alice Lau and Ornstein, {David K.}",
year = "2007",
month = "11",
day = "1",
doi = "10.1002/pros.20628",
language = "English",
volume = "67",
pages = "1601--1613",
journal = "Prostate",
issn = "0270-4137",
publisher = "Wiley-Liss Inc.",
number = "15",

}

TY - JOUR

T1 - Culture requirements of prostatic epithelial cell lines for acinar morphogenesis and lumen formation in vitro

T2 - Role of extracellular calcium

AU - Tyson, Darren R.

AU - Inokuchi, Junichi

AU - Tsunoda, Toshiyuki

AU - Lau, Alice

AU - Ornstein, David K.

PY - 2007/11/1

Y1 - 2007/11/1

N2 - BACKGROUND. Three-dimensional (3D) culture of benign prostatic epithelial cell lines can recapitulate acinar morphogenesis in vitro, but the broad applicability of this approach has not been described. The present studies examine the culture conditions important for prostatic acinar morphogenesis in vitro and the role of extracellular calcium in this process. METHODS. With optimized culture conditions, RWPE-1, pRNS-1-1, PZ-HPV-7, PNT1A, BPH-1, and PrEC were analyzed for their ability to undergo acinar morphogenesis in 3D culture and by immunoblotting. RWPE-1 cells were further examined for the effects of calcium on morphology, E-cadherin membrane localization and multicellular layering in 2D culture and for acinar morphogenesis, luminal apoptosis, and luminal filling in 3D. RESULTS. Cell lines grown in low-calcium medium have the ability to form acinar structures with lumens, which correlates with E-cadherin expression, but low calcium is not required for this process. Adding CaCl 2 to the medium strongly inhibits lumen formation, luminal apoptosis and induces luminal filling, and luminal filling is blocked by an interfering antibody. CONCLUSIONS. Optimized medium composition allows nearly all seeded RWPE-1 cells to undergo acinar morphogenesis, forming consistent structures representative of normal adult prostate glands. Low-calcium-containing medium appears selective for cells capable of undergoing acinar morphogenesis in vitro, and branching and luminal space within the acini are strongly influenced by extracellular calcium levels, likely through the actions of E-cadherin. These results provide important information about a relevant in vitro model with which to study prostate development and carcinogenesis and highlight the importance of extracellular calcium in regulating 3D morphology.

AB - BACKGROUND. Three-dimensional (3D) culture of benign prostatic epithelial cell lines can recapitulate acinar morphogenesis in vitro, but the broad applicability of this approach has not been described. The present studies examine the culture conditions important for prostatic acinar morphogenesis in vitro and the role of extracellular calcium in this process. METHODS. With optimized culture conditions, RWPE-1, pRNS-1-1, PZ-HPV-7, PNT1A, BPH-1, and PrEC were analyzed for their ability to undergo acinar morphogenesis in 3D culture and by immunoblotting. RWPE-1 cells were further examined for the effects of calcium on morphology, E-cadherin membrane localization and multicellular layering in 2D culture and for acinar morphogenesis, luminal apoptosis, and luminal filling in 3D. RESULTS. Cell lines grown in low-calcium medium have the ability to form acinar structures with lumens, which correlates with E-cadherin expression, but low calcium is not required for this process. Adding CaCl 2 to the medium strongly inhibits lumen formation, luminal apoptosis and induces luminal filling, and luminal filling is blocked by an interfering antibody. CONCLUSIONS. Optimized medium composition allows nearly all seeded RWPE-1 cells to undergo acinar morphogenesis, forming consistent structures representative of normal adult prostate glands. Low-calcium-containing medium appears selective for cells capable of undergoing acinar morphogenesis in vitro, and branching and luminal space within the acini are strongly influenced by extracellular calcium levels, likely through the actions of E-cadherin. These results provide important information about a relevant in vitro model with which to study prostate development and carcinogenesis and highlight the importance of extracellular calcium in regulating 3D morphology.

UR - http://www.scopus.com/inward/record.url?scp=35648948933&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=35648948933&partnerID=8YFLogxK

U2 - 10.1002/pros.20628

DO - 10.1002/pros.20628

M3 - Article

C2 - 17705248

AN - SCOPUS:35648948933

VL - 67

SP - 1601

EP - 1613

JO - Prostate

JF - Prostate

SN - 0270-4137

IS - 15

ER -