TY - JOUR
T1 - Protein ligand and nanotopography separately drive the phenotype of mouse embryonic stem cells
AU - Ghorbani, Sadegh
AU - Christine Füchtbauer, Annette
AU - Møllebjerg, Andreas
AU - Møller Martensen, Pia
AU - Hvidbjerg Laursen, Sara
AU - Christian Evar Kraft, David
AU - Kjems, Jørgen
AU - Meyer, Rikke Louise
AU - Rahimi, Karim
AU - Foss, Morten
AU - Füchtbauer, Ernst Martin
AU - Sutherland, Duncan S.
N1 - Publisher Copyright:
© 2023
PY - 2023/10
Y1 - 2023/10
N2 - Biochemical and biomechanical signals regulate stem cell function in the niche environments in vivo. Current in vitro culture of mouse embryonic stem cells (mESC) uses laminin (LN-511) to provide mimetic biochemical signaling (LN-521 for human systems) to maintain stemness. Alternative approaches propose topographical cues to provide biomechanical cues, however combined biochemical and topographic cues may better mimic the in vivo environment, but are largely unexplored for in vitro stem cell expansion. In this study, we directly compare in vitro signals from LN-511 and/or topographic cues to maintain stemness, using systematically-varied submicron pillar patterns or flat surfaces with or without preadsorbed LN-511. The adhesion of cells, colony formation, expression of the pluripotency marker,octamer-binding transcription factor 4 (Oct4), and transcriptome profiling were characterized. We observed that either biochemical or topographic signals could maintain stemness of mESCs in feeder-free conditions, indicated by high-level Oct4 and gene profiling by RNAseq. The combination of LN-511 with nanotopography reduced colony growth, while maintaining stemness markers, shifted the cellular phenotype indicating that the integration of biochemical and topographic signals is antagonistic. Overall, significantly faster (up to 2.5 times) colony growth was observed at nanotopographies without LN-511, suggesting for improved ESC expansion.
AB - Biochemical and biomechanical signals regulate stem cell function in the niche environments in vivo. Current in vitro culture of mouse embryonic stem cells (mESC) uses laminin (LN-511) to provide mimetic biochemical signaling (LN-521 for human systems) to maintain stemness. Alternative approaches propose topographical cues to provide biomechanical cues, however combined biochemical and topographic cues may better mimic the in vivo environment, but are largely unexplored for in vitro stem cell expansion. In this study, we directly compare in vitro signals from LN-511 and/or topographic cues to maintain stemness, using systematically-varied submicron pillar patterns or flat surfaces with or without preadsorbed LN-511. The adhesion of cells, colony formation, expression of the pluripotency marker,octamer-binding transcription factor 4 (Oct4), and transcriptome profiling were characterized. We observed that either biochemical or topographic signals could maintain stemness of mESCs in feeder-free conditions, indicated by high-level Oct4 and gene profiling by RNAseq. The combination of LN-511 with nanotopography reduced colony growth, while maintaining stemness markers, shifted the cellular phenotype indicating that the integration of biochemical and topographic signals is antagonistic. Overall, significantly faster (up to 2.5 times) colony growth was observed at nanotopographies without LN-511, suggesting for improved ESC expansion.
KW - Biochemical cues
KW - Biomechanical signal
KW - Embryonic stem cells
KW - Nanotopography
UR - http://www.scopus.com/inward/record.url?scp=85165090857&partnerID=8YFLogxK
U2 - 10.1016/j.biomaterials.2023.122244
DO - 10.1016/j.biomaterials.2023.122244
M3 - Journal article
C2 - 37459700
AN - SCOPUS:85165090857
SN - 0142-9612
VL - 301
JO - Biomaterials
JF - Biomaterials
M1 - 122244
ER -