Genetically encoded Ca2+ -sensor reveals details of porcine endothelial cell activation upon contact with human serum.

Wuensch, Annegret; Kameritsch, Petra; Sfriso, Riccardo; Jemiller, Eva-Maria; Bähr, Andrea; Kurome, Mayuko; Kessler, Barbara; Kemter, Elisabeth; Kupatt, Christian; Reichart, Bruno; Rieben, Robert; Wolf, Eckhard; Klymiuk, Nikolai (2020). Genetically encoded Ca2+ -sensor reveals details of porcine endothelial cell activation upon contact with human serum. Xenotransplantation, 27(5), e12585. Wiley 10.1111/xen.12585

[img]
Preview
Text
xen.12585.pdf - Published Version
Available under License Creative Commons: Attribution (CC-BY).

Download (1MB) | Preview

The activation of the endothelial surface in xenografts is still a poorly understood process and the consequences are unpredictable. The role of Ca2+ -messaging during the activation of endothelial cells is well recognized and routinely measured by synthetic Ca2+ -sensitive fluorophors. However, these compounds require fresh loading immediately before each experiment and in particular when grown in state-of-the-art 3D cell culture systems, endothelial cells are difficult to access with such sensors. Therefore, we developed transgenic pigs expressing a Ca2+ -sensitive protein and examined its principal characteristics. Primary transgenic endothelial cells stimulated by ATP showed a definite and short influx of Ca2+ into the cytosol, whereas exposure to human serum resulted in a more intense and sustained response. Surprisingly, not all endothelial cells reacted identically to a stimulus, rather activation took place in adjacent cells in a timely decelerated way and with distinct intensities. This effect was again more pronounced when cells were stimulated with human serum. Finally, we show clear evidence that antibody binding alone significantly activated endothelial cells, whereas antibody depletion dramatically reduced the stimulatory potential of serum. Transgenic porcine endothelial cells expressing a Ca2+ -sensor represent an interesting tool to dissect factors inducing activation of porcine endothelial cells after exposure to human blood or serum.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Mu50 > Forschungsgruppe Herz und Gefässe
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Mu50 > Forschungsgruppe Handchirurgie
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Mu50

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Sfriso, Riccardo and Rieben, Robert

Subjects:

600 Technology > 610 Medicine & health

ISSN:

0908-665X

Publisher:

Wiley

Language:

English

Submitter:

Marla Rittiner

Date Deposited:

17 Aug 2020 18:25

Last Modified:

04 Oct 2020 01:33

Publisher DOI:

10.1111/xen.12585

PubMed ID:

32056300

Uncontrolled Keywords:

antibody depletion complement activation genetically encoded Ca2+-sensor porcine endothelial cell activation

BORIS DOI:

10.7892/boris.145962

URI:

https://boris.unibe.ch/id/eprint/145962

Actions (login required)

Edit item Edit item
Provide Feedback