Ek0416

The total protein content of the conditioned medium obtained from MSCs in 2D and 3D cultures was measured by the bicinchoninic acid (BCA) method, a colorimetric method. The procedure was performed with the directions in the commercially purchased protein quantification kit (KTD3001, Abbkine Co., USA), the absorbances were measured with a microplate reader (Allsheng AMR-100 Microplate Reader AS-16050-00), and the protein amount was calculated as μg/ml. For the cytokine analysis in the conditioned medium, commercially purchased “enzyme-linked immunosorbent assay” (ELISA) kits were used, and the levels of IFN-γ (EK0373, Boster, Wuhan, China), IL 17 (EK0430, Boster, Wuhan, China), IL-4 (EK0404, Boster, Wuhan, China), and IL-10 (EK0416, Boster, Wuhan, China) were detected. The manufacturer’s instructions were followed during the analyses, and the measurements were made with a 450-nm microplate reader (Allsheng AMR-100 Microplate Reader AS-16050-00). Each sample was performed in duplicates to reduce the error. The concentrations of the samples were calculated as pg/ml.

The IL-6, IL-7, IL-10, IFN-γ, TGF-β and anti-AchR antibodies levels in the sera of the MG patients and NCs were measured via ELISA Kits (EK0410, EK0779, EK0416, EK0373, EK0513 and ElisaRSR^TM AChRAb, respectively, from Boster Biological Technology, Pleasanton, CA, USA), in accordance with manufacturer instructions. In particular, the cut-off value for anti-AchR antibody positive was 0.45 nmol/L.

The concentration levels of IL-1β, IL-6, IL-10 and TNF-α in the cell culture supernatants were measured using ELISA kits (EK0392, EK0410, EK0416, EK0525, Boster Biological Technology, China) according to the manufacturers’ protocols.