3D Tissue Explant and Single-Cell Suspension Organoid Culture Systems for Ex Vivo Drug Testing on Human Tonsil-Derived T Follicular Helper Cells

2021 ◽  
pp. 267-288
Author(s):  
Angelika Schmidt ◽  
Dirk Baumjohann
Author(s):  
Debby A. Jennings ◽  
Michael J. Morykwas ◽  
Louis C. Argenta

Grafts of cultured allogenic or autogenic keratlnocytes have proven to be an effective treatment of chronic wounds and burns. This study utilized a collagen substrate for keratinocyte and fibroblast attachment. The substrate provided mechanical stability and augmented graft manipulation onto the wound bed. Graft integrity was confirmed by light and transmission electron microscopy.Bovine Type I dermal collagen sheets (100 μm thick) were crosslinked with 254 nm UV light (13.5 Joules/cm2) to improve mechanical properties and reduce degradation. A single cell suspension of third passage neonatal foreskin fibroblasts were plated onto the collagen. Five days later, a single cell suspension of first passage neonatal foreskin keratinocytes were plated on the opposite side of the collagen. The grafts were cultured for one month.The grafts were fixed in phosphate buffered 4% formaldehyde/1% glutaraldehyde for 24 hours. Graft pieces were then washed in 0.13 M phosphate buffer, post-fixed in 1% osmium tetroxide, dehydrated, and embedded in Polybed 812.


protocols.io ◽  
2021 ◽  
Author(s):  
Sara Napolitano ◽  
Michal Slyper ◽  
Sébastien Vigneau ◽  
Asaf Rotem ◽  
Bruce Johnson ◽  
...  

2021 ◽  
Author(s):  
Steven B. Wells ◽  
Peter A. Szabo ◽  
Basak Ural ◽  
Maya M.L. Poon

This protocol describes a method for the isolation of the immune cells, structural and epithelial cells, and progenitors from human lung sections of about two grams. By providing defined media formulations, volumes at each step, and a defined dilution factor for density centrifugation, it yields consistent single-cell suspensions across samples.


MethodsX ◽  
2018 ◽  
Vol 5 ◽  
pp. 1287-1290 ◽  
Author(s):  
Erica Bresciani ◽  
Elizabeth Broadbridge ◽  
Paul P. Liu

protocols.io ◽  
2021 ◽  
Author(s):  
Isaac Wakiro ◽  
Sara Napolitano ◽  
Sébastien Vigneau ◽  
Asaf Rotem ◽  
Orit Rozenblatt-Rosen ◽  
...  

2016 ◽  
pp. 101-112 ◽  
Author(s):  
Valentina Benedetti ◽  
Valerio Brizi ◽  
Christodoulos Xinaris

2015 ◽  
Vol 272 (12) ◽  
pp. 3727-3734 ◽  
Author(s):  
Aleksandra Barac ◽  
Marina Pekmezovic ◽  
Vesna Tomic Spiric ◽  
Aleksandar Trivic ◽  
Jelena Marinkovic ◽  
...  

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