Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder Cells

Human embryonic stem (hES) cells represent an important tool to study early cell development. The previously described use of human recombinant laminin (LN) 521 represented a step forward in generating clinically safe culture conditions. To test the short-term effect of LN521 on cultured hES cells,...

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Main Authors: Halima Albalushi, Magdalena Kurek, Leif Karlsson, Luise Landreh, Kristín Rós Kjartansdóttir, Olle Söder, Outi Hovatta, Jan-Bernd Stukenborg
Format: Article
Language:English
Published: Hindawi Limited 2018-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2018/7127042
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spelling doaj-6452a9331bd54054b0e10cdbfab014112020-11-24T22:49:49ZengHindawi LimitedStem Cells International1687-966X1687-96782018-01-01201810.1155/2018/71270427127042Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder CellsHalima Albalushi0Magdalena Kurek1Leif Karlsson2Luise Landreh3Kristín Rós Kjartansdóttir4Olle Söder5Outi Hovatta6Jan-Bernd Stukenborg7Nordfertil Research Lab Stockholm, Department of Women’s and Children’s Health, Karolinska Institutet and University Hospital Karolinska Institutet, Stockholm, SwedenNordfertil Research Lab Stockholm, Department of Women’s and Children’s Health, Karolinska Institutet and University Hospital Karolinska Institutet, Stockholm, SwedenPaediatric Endocrinology Unit, Department of Women’s and Children’s Health, Karolinska Institutet and University Hospital Karolinska Institutet, Stockholm, SwedenNordfertil Research Lab Stockholm, Department of Women’s and Children’s Health, Karolinska Institutet and University Hospital Karolinska Institutet, Stockholm, SwedenPaediatric Endocrinology Unit, Department of Women’s and Children’s Health, Karolinska Institutet and University Hospital Karolinska Institutet, Stockholm, SwedenPaediatric Endocrinology Unit, Department of Women’s and Children’s Health, Karolinska Institutet and University Hospital Karolinska Institutet, Stockholm, SwedenDepartment of Clinical Science, Intervention and Technology (CLINTEC), Karolinska Institutet and University Hospital Karolinska Institutet, Stockholm, SwedenNordfertil Research Lab Stockholm, Department of Women’s and Children’s Health, Karolinska Institutet and University Hospital Karolinska Institutet, Stockholm, SwedenHuman embryonic stem (hES) cells represent an important tool to study early cell development. The previously described use of human recombinant laminin (LN) 521 represented a step forward in generating clinically safe culture conditions. To test the short-term effect of LN521 on cultured hES cells, five male hES cell lines were cultured on human foreskin fibroblasts (hFFs), Matrigel, LN521, and LN121 and characterized by qPCR, immunofluorescence analysis, as well as their potential for three-germ layer differentiation. Variations in gene expression related to pluripotency, stemness, and testicular cells at different passages and culture conditions were evaluated by qPCR. All cell lines expressed pluripotency markers at protein and RNA level and were able to differentiate into cell types of the three germ layers after being cultured on LN521 for nine passages. Reduction in variation of pluripotency marker expression could be observed after culturing the cells on LN521 for nine passages. hES cells cultured on LN521 exhibited less differentiation, faster cell growth, and attachment when compared to hES cells cultured on LN121 or Matrigel. Our results indicate a positive effect of LN521 in stabilizing pluripotency gene expression and might be the first step towards more controllable and robust culture conditions for hES cells.http://dx.doi.org/10.1155/2018/7127042
collection DOAJ
language English
format Article
sources DOAJ
author Halima Albalushi
Magdalena Kurek
Leif Karlsson
Luise Landreh
Kristín Rós Kjartansdóttir
Olle Söder
Outi Hovatta
Jan-Bernd Stukenborg
spellingShingle Halima Albalushi
Magdalena Kurek
Leif Karlsson
Luise Landreh
Kristín Rós Kjartansdóttir
Olle Söder
Outi Hovatta
Jan-Bernd Stukenborg
Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder Cells
Stem Cells International
author_facet Halima Albalushi
Magdalena Kurek
Leif Karlsson
Luise Landreh
Kristín Rós Kjartansdóttir
Olle Söder
Outi Hovatta
Jan-Bernd Stukenborg
author_sort Halima Albalushi
title Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder Cells
title_short Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder Cells
title_full Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder Cells
title_fullStr Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder Cells
title_full_unstemmed Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder Cells
title_sort laminin 521 stabilizes the pluripotency expression pattern of human embryonic stem cells initially derived on feeder cells
publisher Hindawi Limited
series Stem Cells International
issn 1687-966X
1687-9678
publishDate 2018-01-01
description Human embryonic stem (hES) cells represent an important tool to study early cell development. The previously described use of human recombinant laminin (LN) 521 represented a step forward in generating clinically safe culture conditions. To test the short-term effect of LN521 on cultured hES cells, five male hES cell lines were cultured on human foreskin fibroblasts (hFFs), Matrigel, LN521, and LN121 and characterized by qPCR, immunofluorescence analysis, as well as their potential for three-germ layer differentiation. Variations in gene expression related to pluripotency, stemness, and testicular cells at different passages and culture conditions were evaluated by qPCR. All cell lines expressed pluripotency markers at protein and RNA level and were able to differentiate into cell types of the three germ layers after being cultured on LN521 for nine passages. Reduction in variation of pluripotency marker expression could be observed after culturing the cells on LN521 for nine passages. hES cells cultured on LN521 exhibited less differentiation, faster cell growth, and attachment when compared to hES cells cultured on LN121 or Matrigel. Our results indicate a positive effect of LN521 in stabilizing pluripotency gene expression and might be the first step towards more controllable and robust culture conditions for hES cells.
url http://dx.doi.org/10.1155/2018/7127042
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