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Isolation and Time Lapse Microscopy of Highly Pure Hepatic Stellate Cells
Joint Authors
Heymann, Felix
Weiskirchen, Ralf
Bartneck, Matthias
Warzecha, Klaudia Theresa
Tag, Carmen Gabriele
Sauer-Lehnen, Sibille
Trautwein, Christian
Tacke, Frank
Source
Issue
Vol. 2015, Issue 2015 (31 Dec. 2015), pp.1-13, 13 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2015-07-16
Country of Publication
Egypt
No. of Pages
13
Main Subjects
Abstract EN
Hepatic stellate cells (HSC) are the main effector cells for liver fibrosis.
We aimed at optimizing HSC isolation by an additional step of fluorescence-activated cell sorting (FACS) via a UV laser.
HSC were isolated from livers of healthy mice and animals subjected to experimental fibrosis.
HSC isolation by iohexol- (Nycodenz) based density centrifugation was compared to a method with subsequent FACS-based sorting.
We assessed cellular purity, viability, morphology, and functional properties like proliferation, migration, activation marker, and collagen expression.
FACS-augmented isolation resulted in a significantly increased purity of stellate cells (>99%) compared to iohexol-based density centrifugation (60–95%), primarily by excluding doublets of HSC and Kupffer cells (KC).
Importantly, this method is also applicable to young animals and mice with liver fibrosis.
Viability, migratory properties, and HSC transdifferentiation in vitro were preserved upon FACS-based isolation, as assessed using time lapse microscopy.
During maturation of HSC in culture, we did not observe HSC cell division using time lapse microscopy.
Strikingly, FACS-isolated, differentiated HSC showed very limited molecular and functional responses to LPS stimulation.
In conclusion, isolating HSC from mouse liver by additional FACS significantly increases cell purity by removing contaminations from other cell populations especially KC, without affecting HSC viability, migration, or differentiation.
American Psychological Association (APA)
Bartneck, Matthias& Warzecha, Klaudia Theresa& Tag, Carmen Gabriele& Sauer-Lehnen, Sibille& Heymann, Felix& Trautwein, Christian…[et al.]. 2015. Isolation and Time Lapse Microscopy of Highly Pure Hepatic Stellate Cells. Analytical Cellular Pathology،Vol. 2015, no. 2015, pp.1-13.
https://search.emarefa.net/detail/BIM-1052353
Modern Language Association (MLA)
Bartneck, Matthias…[et al.]. Isolation and Time Lapse Microscopy of Highly Pure Hepatic Stellate Cells. Analytical Cellular Pathology No. 2015 (2015), pp.1-13.
https://search.emarefa.net/detail/BIM-1052353
American Medical Association (AMA)
Bartneck, Matthias& Warzecha, Klaudia Theresa& Tag, Carmen Gabriele& Sauer-Lehnen, Sibille& Heymann, Felix& Trautwein, Christian…[et al.]. Isolation and Time Lapse Microscopy of Highly Pure Hepatic Stellate Cells. Analytical Cellular Pathology. 2015. Vol. 2015, no. 2015, pp.1-13.
https://search.emarefa.net/detail/BIM-1052353
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1052353