Tissue preservation solution

This work was reviewed and approved by the Institutional Review Boards of Nantong University Hospital (2020-K013). Parents of study participants signed an Informed consent form. For scRNA-seq, TMJC tissue was isolated by surgical excision. The entire mandible, including the mandibular body and TMJ (condyle, joint capsule, joint disc, part of fibrous ligament and temporo-articular fossa), was isolated along the mandibular labiobuccal migration. The mandibular body, condyle and coracoid process were separated from the rest of the TMJ. The condyle is separated from the mandibular body through the neck of the condyle (Additional file 1: Fig. S1a). The left condyle was cut into pieces as far as possible and placed in tissue preservation solution (Singleron Biotechnologies, Nanjing, China), and then transported to the laboratory by cold chain for cell dissociation and single cell sequencing.

Fresh breast tissues were stored in Tissue Preservation Solution (Singleron Biotechnologies, Nanjing, China) and placed on ice after the biopsy within 30 min. The specimens were washed three times and then cut into slices of 1 to 2 mm. Subsequently, the tissue pieces were digested in a 15 ml centrifuge tube at 37 °C with continuous agitation for 15 min. Then, they were centrifuged at 500 × g for 5 min and suspended gently with PBS (HyClone, USA). Finally, the samples were stained with trypan blue (Sigma, USA), and cell viability was evaluated under a phase-contrast microscope (Nikon, Japan).