For histology and immunohistochemistry (IHC), unfixed chondrocyte‐laden alginate beads were covered in Tissue‐Tek (Thermo Fisher), snap frozen in liquid nitrogen, and cut in 14–20 μm sections at −20°C using a cryostat. Sections were mounted on Superfrost Plus Gold microscopic slides (Thermo Scientific) and stained with Alcian Blue (pH 1.0) for histological examination or processed for IHC. For IHC, sections were incubated for 2 h with monoclonal antibodies against type II collagen (II‐II6B3, Developmental Studies Hybridoma Bank) or type VI collagen (MAB3303, EMD Millipore). Following incubation, sections were washed in PBS and incubated for 30 min with EnVision (Dako). Finally, sections were incubated with AEC substrate for 10 min and visualized using microscopy. Postprocessing was undertaken using NIS Elements software (Nikon Instruments Europe B.V.).
Control cartilage samples derived from goat ears were embedded in paraffin using standard histological techniques, cut in 5 μm sections, and stained using Alcian Blue, Mayer's Hematoxylin‐Eosin, type II and type VI collagen to assess glycosaminoglycan distribution, morphology, and collagen distribution, respectively (Supporting Information Fig.S1 ).
Control cartilage samples derived from goat ears were embedded in paraffin using standard histological techniques, cut in 5 μm sections, and stained using Alcian Blue, Mayer's Hematoxylin‐Eosin, type II and type VI collagen to assess glycosaminoglycan distribution, morphology, and collagen distribution, respectively (Supporting Information Fig.