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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Online Access: | http://dx.doi.org/10.1155/2018/7127042 |
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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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