JoVE 비디오를 활용하시려면 도서관을 통한 기관 구독이 필요합니다. 전체 비디오를 보시려면 로그인하거나 무료 트라이얼을 시작하세요.
An In Vitro Assay for Evaluating the Immunomodulatory Impact of Monocytes on Leukocyte Proliferation
내레이션 대본
To perform an effector suppression assay, begin by setting up an MSC-PBMC co-culture with 2.5 x 106 PBMCs, co-cultured with 250,000 MSCs to ensure enough MSC co-cultured PBMCs for subpopulation selection.
After 48 to 72 hours at 37 degrees Celsius, select the MSC-induced immunomodulatory leukocytes of interest with the appropriate magnetic beads. Then, add CFSE-labeled allogeneic CD4-positive T cells in 1 milliliter of complete leukocyte medium per well, at various experimental ratios.
Next, add anti-CD3/28-conjugated microbeads at a 1-to-1 bead-to-cell ratio to stimulate the CD4-positive T cells. On the third day of co-culture, assess the proliferation of the CFSE-labeled CD4-positive T cells by flow cytometry.
