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Merck

3D-Embedded Cell Cultures to Study Tendon Biology.

Methods in molecular biology (Clifton, N.J.) (2019-02-09)
Renate Gehwolf, Gabriel Spitzer, Andrea Wagner, Christine Lehner, Nadja Weissenbacher, Herbert Tempfer, Andreas Traweger
초록

Tendons harbor various cell populations, including cells displaying classical adult mesenchymal stromal cell criteria. Previous studies have shown that a tenogenic phenotype is more effectively maintained in a 3D cell culture model under mechanical load. This chapter describes a method to isolate tendon-derived cells from rat Achilles tendons and the subsequent formation of 3D-embedded cell cultures. These tendon-like constructs can then be analyzed by various means, including histology, immunohistochemistry, qPCR, or standard protein analysis techniques.

MATERIALS
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Sigma-Aldrich
Aprotinin from bovine lung, BioUltra, 3-8 TIU/mg solid, ≥98% (SDS-PAGE)
Roche
DAPI, 4′,6-Diamidine-2′-phenylindole dihydrochloride