open access

Vol 52, No 3 (2014)
ORIGINAL PAPERS
Published online: 2014-10-10
Submitted: 2014-07-07
Accepted: 2014-09-24
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Growth factors and their receptors derived from human amniotic cells in vitro

Zofia Grzywocz, Ewa Pius-Sadowska, Patrycja Klos, Marek Gryzik, Danuta Wasilewska, Barbara Aleksandrowicz, Malgorzata Dworczynska, Stanislawa Sabalinska, Grazyna Hoser, Boguslaw Machalinski, Jerzy Kawiak
DOI: 10.5603/FHC.2014.0019
·
Folia Histochem Cytobiol 2014;52(3):163-170.

open access

Vol 52, No 3 (2014)
ORIGINAL PAPERS
Published online: 2014-10-10
Submitted: 2014-07-07
Accepted: 2014-09-24

Abstract

In vitro studies have shown that amnion-produced growth factors participated in angiogenesis, re-epithelialization, and immunomodulation. The aim of our study was to investigate the growth factors and receptors produced by human amnion tissue and amniotic cells. Human amnions (hAM) were isolated, and amnion circles were dissected for in vitro analysis. Some amnion fragments were digested by the use of different methods to obtain two cell fractions, which were analysed for mesenchymal and epithelial cell markers. Amniotic circles and human amniotic cell fractions were cultured in a protein-free medium. Proteins secreted into the culture medium were analysed with a human growth factor antibody array. Conditioned culture media were added to human umbilical vein epithelial cells (HUVECs) to test for stimulation of migration (scratch test) and proliferation (Ki67 expression). Fraction 1 cells expressed both cytokeratin and mesenchymal cell markers which indicated that it was composed of a mixture of human amnion epithelial cells (hAECs) and mesenchymal stromal cells (hAMSCs). Fraction 2 cells mainly expressed cytokeratin and, therefore, were designed as hAECs. Secretion of proteins by the cultured cells increased with time. The hAM cultures secreted EGF-R, IGF, and IGFBP-2,-3 and -6; Cell Fraction 1 secreted NT-4, whereas Cell Fraction 2 secreted G-CSF, M-CSF, and PDGF. Conditioned media of hAM cultures stimulated HUVECs migration. We have showed for the first time that human amnions and amniotic cells secreted IGFBP-6, MCSF-R, PDGF-AB, FGF-6, IGFBP-4, NT-4, and VEGF-R3. We found that Cell Fraction 1, Cell Fraction 2, and the whole amnion secreted different proteins, possibly due to different proportions of amnion-derived cells and different cell-cell interactions. The hAM cell factors remained functional in vitro and induced intensified migration of HUVECs. The growth factors and receptors found in amnion or amniotic cell media might be used for regenerative medicine.

Abstract

In vitro studies have shown that amnion-produced growth factors participated in angiogenesis, re-epithelialization, and immunomodulation. The aim of our study was to investigate the growth factors and receptors produced by human amnion tissue and amniotic cells. Human amnions (hAM) were isolated, and amnion circles were dissected for in vitro analysis. Some amnion fragments were digested by the use of different methods to obtain two cell fractions, which were analysed for mesenchymal and epithelial cell markers. Amniotic circles and human amniotic cell fractions were cultured in a protein-free medium. Proteins secreted into the culture medium were analysed with a human growth factor antibody array. Conditioned culture media were added to human umbilical vein epithelial cells (HUVECs) to test for stimulation of migration (scratch test) and proliferation (Ki67 expression). Fraction 1 cells expressed both cytokeratin and mesenchymal cell markers which indicated that it was composed of a mixture of human amnion epithelial cells (hAECs) and mesenchymal stromal cells (hAMSCs). Fraction 2 cells mainly expressed cytokeratin and, therefore, were designed as hAECs. Secretion of proteins by the cultured cells increased with time. The hAM cultures secreted EGF-R, IGF, and IGFBP-2,-3 and -6; Cell Fraction 1 secreted NT-4, whereas Cell Fraction 2 secreted G-CSF, M-CSF, and PDGF. Conditioned media of hAM cultures stimulated HUVECs migration. We have showed for the first time that human amnions and amniotic cells secreted IGFBP-6, MCSF-R, PDGF-AB, FGF-6, IGFBP-4, NT-4, and VEGF-R3. We found that Cell Fraction 1, Cell Fraction 2, and the whole amnion secreted different proteins, possibly due to different proportions of amnion-derived cells and different cell-cell interactions. The hAM cell factors remained functional in vitro and induced intensified migration of HUVECs. The growth factors and receptors found in amnion or amniotic cell media might be used for regenerative medicine.

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Keywords

human amnion; amniotic epithelial cells; amniotic mesenchymal stromal cells; cell culture; growth factors

About this article
Title

Growth factors and their receptors derived from human amniotic cells in vitro

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 52, No 3 (2014)

Pages

163-170

Published online

2014-10-10

DOI

10.5603/FHC.2014.0019

Bibliographic record

Folia Histochem Cytobiol 2014;52(3):163-170.

Keywords

human amnion
amniotic epithelial cells
amniotic mesenchymal stromal cells
cell culture
growth factors

Authors

Zofia Grzywocz
Ewa Pius-Sadowska
Patrycja Klos
Marek Gryzik
Danuta Wasilewska
Barbara Aleksandrowicz
Malgorzata Dworczynska
Stanislawa Sabalinska
Grazyna Hoser
Boguslaw Machalinski
Jerzy Kawiak

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