Human embryonic (hESCs) and induced pluripotent stem cell (hiPSCs) facility

Vascular Organoids
Retinal Organoids

Vascular organoid generated from vascular smooth muscle cells (red) and endothelial cells (green) both differentiated from human embryonic stem cells. Nuclei are shown in blue. Murphy-Fotsis lab

We have established a facility which handles hESCs and hiPSCs. These cells are used for basic research and also as disease models. Projects that the stem cell facility can perform in collaboration and/or under subcontracting include:

Generation of patient specific human induced pluripotent stem cells
CRISPR genome editing of hESCS and hiPSCs
Expression of genes of interest in hESCs and hiPSCs
Differentiation of hESCs/hiPSCs to endothelial and mural cells
Targeted differentiation of patient derived hiPSCs
Formation of gastruloids
Generation of Vascular organoids
Generation of Retinal and other organoids
Vascularisation of organoids
Generation of anterior cruciate ligament
Drug and small molecule semi-high throughput screening of patient derived hiPSCs
Toxicology in human pluripotent and differentiated cell types
Endothelial barrier models to assess drug availability
Advanced imaging techniques

The facility harbours 2 incubators dedicated for work with these cell types, a dissection hood and a biosafety flow cabinet. A stereo microscope for isolation of clones and a Zeiss microscope for visualization and imaging are also available. Stem cells are handled separately from other cell types and liquid nitrogen storage containers harbouring cell stocks are also dedicated to stem cells to avoid potential contamination.

Dissection hood containing a stereo microscope and Zeiss imaging system	A Biosafety cabinet is dedicated to human stem cell work

Relevant Publications
Kostopoulou N, Bellou S, Bagli E, Markou M, Kostaras E, Hyvönen M, Kalaidzidis Y, Angelos Papadopoulos A, Chalmantzi V, Kyrkou A, Panopoulou E, Fotsis T, Murphy C: Embryonic Stem Cells Are Devoid of Macropinocytosis, a Trafficking Pathway for Activin A in Differentiated Cells. J. Cell Sci. 2021 Jul 1;134(13):jcs246892. doi: 10.1242/jcs.246892. Epub 2021 Jul 12.
https://pubmed.ncbi.nlm.nih.gov/34313314/

Chalmantzi V, Simitzi C, Papadopoulos A, Bagli E, Murphy C, Stratakis E, Fotsis T: Culturing human pluripotent stem cells on micropatterned silicon surfaces. In: Methods in Molecular Biology. Springer, New York, NY.  doi.org/10.1007/7651_2021_428. Dec. 2021.

Markou M, Kouroupis D, Fotsis T, Bagli E,  Murphy C. Vascularisation in 3D cell culture. Basic Concepts on 3D Cell culture. Springer. ISBN 978-3-030-66749-8. 2021.
https://doi.org/10.1007/978-3-030-66749-8

Papadopoulos A, Chalmantzi V, Mikhaylichenko O, Hyvönen M, Stellas D, Kanhere A, Heath J, Cunningham DL, Fotsis T, Murphy C: Supporting data on combined transcriptomics and phosphoproteomic analysis of BMP4 signaling in human embryonic stem cells. Data in Brief 35 (2021) 106844.
https://www.sciencedirect.com/science/article/pii/S2352340921001281

Papadopoulos A, Chalmantzi V, Mikhaylichenko O, Hyvönen M, Stellas D, Kanhere A, Heath J, Cunningham DL, Fotsis T, Murphy C: Combined transcriptomics and phosphoproteomic analysis of BMP4 signaling in human embryonic stem cells. Stem Cell Res 50 (2021) 102133.
https://pubmed.ncbi.nlm.nih.gov/33383406/

Markou M, Kouroupis D, Badounas F, Katsouras A, Kyrkou A, Fotsis T, Murphy C*, Bagli E*. Tissue engineering using vascular organoids from human pluripotent stem cell derived mural cell phenotypes. Front Bioeng Biotechnol, section Tissue Engineering and Regenerative Medicine. 8(2020) article 278, 1-20.* joint corresponding authors.
https://pubmed.ncbi.nlm.nih.gov/32363181/

Tsolis K, Bagli E, Kanaki K, Zografou S, Carpentier S, Bei E, Christoforidis S, Zervakis M, Murphy C, Fotsis T, Economou A. Proteome changes during transition from human embryonic to vascular progenitor cells. J Proteome Res 15 (2016) 1995-2007.
https://www.ncbi.nlm.nih.gov/pubmed/27146950

Kyrkou A, Stellas D, Syrrou M, Klinakis A, Fotsis T, Murphy C: Generation of human induced pluripotent stem cells in defined, feeder-free conditions. Stem Cell Res, 17 (2016) 458-460.
https://www.sciencedirect.com/science/article/pii/S1873506116300381

Kouroupis D, Kyrkou A, Triantafyllidi E, Katsimpoulas M, Chalepakis G, Goussia A,  Georgoulis A, Murphy C, Fotsis T: Generation of stem cell-based bioartificial anterior cruciate ligament (ACL) grafts for effective ACL rupture repair. Stem Cell Res 17 (2016) 448-457.
https://www.ncbi.nlm.nih.gov/pubmed/27217303

Contact
Stem cells:
Carol Murphy
Maria Markou
Eleni Bagli
Advanced imaging:
Sofia Bellou