Human Knee Meniscus Collection Protocol for scRNA-seq
molmer, Martin Lotz
Abstract
Meniscus is taken from the medial knee compartment is collected for scRNA-seq processing. The attached image indicates where the meniscal tissue is collected from.
Before start
Knee blocks are shipped on wet ice and received within 48 hours post-mortem from certified tissue banks. The average time from receiving the sample to harvesting cartilage from both knees is about 2 hours. Pictures are taken of knee tissues and macroscopic scoring of the cartilage is completed.
Steps
Tissue Collection for MERFISH
Three pieces of tissue are placed in 12mLs, per region, of
Pour off fixation solution and replace with
Prepare harvesting area with sterile drapes, tools and gauze. All harvesting is completed within an aseptic environment.
Wipe down the knee blocks with 95% ethanol prior to opening the joint capsule.
Once the joint capsule is opened, the femur and tibia are disarticulated and menisci are resected.
A full-thickness (sagittal) piece of medial meniscus is cut out using a #21 scalpel; the piece measures about 1.60cmX15mmx8mm and weighs around 1g. This is collected into a 50mL tube that is filled with 40mLs of
Procurement of Medial Femoral Cartilage With Subchondral Bone for MERFISH: Using an autoclaved 12” hacksaw with 24TPI blade, cut a 3-5mm slab from the middle region of the medial condyle.
