Establishment and characterization of a new human first trimester Trophoblast cell line, AL07.
Cytokeratin 7
Pregnancy
Primary trophoblasts
Trophoblast cell line
hTERT
Journal
Placenta
ISSN: 1532-3102
Titre abrégé: Placenta
Pays: Netherlands
ID NLM: 8006349
Informations de publication
Date de publication:
10 2020
10 2020
Historique:
received:
26
02
2020
revised:
20
07
2020
accepted:
12
08
2020
pubmed:
15
9
2020
medline:
11
8
2021
entrez:
14
9
2020
Statut:
ppublish
Résumé
The limited cell number of primary trophoblasts and contamination of trophoblast cell lines promote us to develop a novel stable trophoblast cell line. Primary trophoblast cells were isolated from first-trimester placenta and telomerase-induced immortalization was used to immortalize these cells. Subsets of cells were then evaluated by flow cytometry using CK7, HLA-G, CD45 and CD14, specific markers for trophoblast cells, extra-villous trophoblast, pan leucocyte and monocyte/macrophage, respectively. Immunofluorescence staining and immunocytochemistry were used to detect CK7 expression in trophoblast cells. The level of secreted human Chorionic Gonadotropin (hCG) was measured by electrochemiluminescence (ECL). The Bio-Plex MAGPIX System was used to analyze the cytokines and chemokines produced by AL07 cell line. We were able to isolate primary trophoblast cells from several first-trimester placentas. One clone, AL07 trophoblast cells, isolated from a week 7 placenta, was morphologically stable and positive for the expression of CK7 by immunofluorescence and immunocytochemistry staining. Characterization of AL07 cells reveled that they are CD45 or CD14 negative and had constitutive secretion of hCG and low HLA-G expression. Furthermore, clone AL07 secret high levels of several cytokines and chemokines, including IL-6, IL-8 and VEGF, and moderately secreted MCP-1 IP-10 and RANTES. We report the successful isolation, immortalization and characterization of AL07 cells, a novel cell clone isolated from first trimester human placenta. The clone is free of contamination of immune cells, and exhibits similar cytokine profile as other trophoblast cell lines. This new cytotrophoblast-like AL07 cell, can be a valuable tool for in-vitro trophoblast studies in the future.
Identifiants
pubmed: 32927240
pii: S0143-4004(20)30266-6
doi: 10.1016/j.placenta.2020.08.013
pmc: PMC8237240
mid: NIHMS1713706
pii:
doi:
Substances chimiques
Chorionic Gonadotropin
0
Cytokines
0
Types de publication
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
122-132Subventions
Organisme : NIAID NIH HHS
ID : R01 AI145829
Pays : United States
Organisme : NCATS NIH HHS
ID : UL1 TR001863
Pays : United States
Informations de copyright
Copyright © 2020 Elsevier Ltd. All rights reserved.
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