Pe mouse anti rat cd8a

The test of IgG, IgM was performed as following. The blood of experimental animals was taken by tail vein cut off to test the immune indexes. Peripheral blood of rats was collected in a 1.5 ml centrifugal tube. In order to prepare serum, peripheral blood of rats was centrifuged for 10 min at 1000 g. The upper serum was collected and stored at −20°C. The serum was placed at room temperature for 30 min before use, then tested by ELISA method according to the operation instructions of ELISA kit. At the end of the immune period (4 and 26 weeks), the rats were euthanized, and the spleens were taken aseptically. Each sample was grinded to prepare a single cell suspension. Cell suspension was collected. RPMI1640 containing 10% FCS was added to adjust the cell concentration to 2 × 10⁶ cells/ml. The above lymphocyte suspensions were stained with FITC mouse anti-rat CD4 (Lot: 5299960) and PE mouse anti-rat CD8a (Lot: 6175730) labeled with BioLegend fluorescence, and then analyzed by flow cytometry.

Biphasic calcium phosphate ceramics particles with 61.64% porosity and 0.8617 ± 0.0068 g/cm³ density (d ≤ 2mm) were purchased from National Engineering Research Center for Biomaterials (Sichuan University, China). Enzyme-linked immunosorbent assay (ELISA) kit for Immunoglobulin G (IgG) and ELISA kit for Immunoglobulin M (IgM) were purchased from Cloud-Clone Corp (USA). Fluorescein isothiocyanate (FITC) mouse Anti-Rat CD4 and PE mouse Anti-Rat CD8a were purchased from BioLegend (USA). Roswell Park Memorial Institute 1640 (RPMI 1640) and fetal bovine serum (FCS) were purchased from HYCLONE (USA).
Experimental rats were purchased from Chengdu Dashuo Experimental Animal Co., Ltd (China). All animal studies were approved by the Sichuan University Medical Ethics Committee. All animal procedures were performed in accordance with the guidelines for care and use of laboratory animals of Sichuan University.