TY - JOUR
T1 - Laminin 521 Stabilizes the Pluripotency Expression Pattern of Human Embryonic Stem Cells Initially Derived on Feeder Cells
AU - Albalushi, Halima
AU - Kurek, Magdalena
AU - Karlsson, Leif
AU - Landreh, Luise
AU - Kjartansdóttir, Kristín Rós
AU - Söder, Olle
AU - Hovatta, Outi
AU - Stukenborg, Jan-Bernd
N1 - Funding Information:
The authors are grateful to Dr. L. Antonsson for her assistance with the cell cultures and to Biolamina for sending test vials of LN121 for their experiment. Karyotyping was performed at Finnish Functional Genomics Centre, University of Turku, Turku, Finland. The authors would also like to gratefully acknowledge the aid of the core facility at Novum (BEA (Bioinformatics and Expression Analysis)), which is supported by the Board of Research of the Karolinska Institutet and the Research Committee of the Karolinska University Hospital. Financial support was received from Sällskåpet Barnavård in Stockholm, the Sigurd and Elsa Goljes Minne Foundation, the Swedish Research Council/Finnish Academy of Science, Kronprinsessan Lovi-sas Förening för Barnasjukvård/Stiftelsen Axel Tielmans Minnesfond, Samariten Foundation, and the Emil och Wera Cornells Foundation. Magdalena Kurek was financially supported by EU-FP7-PEOPLE-2013-ITN 603568 “Grow-sperm”. Halima Albalushi was financially supported by a grant from the Sultan Qaboos University in Oman.
Publisher Copyright:
Copyright © 2018 Halima Albalushi et al.
PY - 2018
Y1 - 2018
N2 - 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.
AB - 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.
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U2 - 10.1155/2018/7127042
DO - 10.1155/2018/7127042
M3 - Article
C2 - 29535778
AN - SCOPUS:85047297273
SN - 1687-966X
VL - 2018
SP - 7127042
JO - Stem Cells International
JF - Stem Cells International
M1 - 7127042
ER -