Lympholyte h cell separation medium

The samples were washed with sterile cold PBS (Gibco, Dublin, Ireland) immediately following surgery. Subcutaneous fat was thoroughly removed. Pericapsule tissues were also removed from capsules by trimming with forceps (Fig. 1). These capsules and pericapsule tissues were used as samples. RPMI 1640 (Invitrogen Life Technologies, Carlsbad, Calif.) containing 10% FBS (Biowest, Nuaillé, France) and Anti-Anti (1:100; Gibco, Dublin, Ireland) without additional exogenous cytokines was utilized as a culture medium. Samples were placed in RPMI 1640 (Invitrogen Life Technologies, Carlsbad, Calif.) containing 200 U/mL collagenase type IV (Worthington, Lakewood, N.J.) and 0.05% DNaseⅠ (Sigma-Aldrich, St. Louis, Mo.) for 120 min at 37°C. To generate single-cell suspensions, these pieces were aspirated with a 50-cm³ syringe up and down 10 times. The sample was then filtered three times through a sterile mesh. In some experiments, debris was excluded using lympholyte-H cell separation medium (Cedarlane, Ontario, Canada) according to the manufacturer’s instructions.