2.2 Model Replication

2.2 Model Replication

Referencing Yao Jun’s improved method [24]:

2.2.1 Preparation of Thymus and Lymph Node Cell Suspension

DBA/2 mice are decapitated, disinfected by soaking in 70% ethanol, fixed with the abdomen facing up, the thoracic cavity is opened, and the thymus along with cervical, submandibular, axillary, inguinal, and mesenteric lymph nodes are removed using surgical scissors. The blood is washed away with saline in a petri dish, and connective tissues are removed. The cells are washed twice in a petri dish containing a small amount of RPMI-1640 culture medium. After gentle grinding, they are filtered through a 200-mesh nylon blood filter to separate individual cells. The cells are then adjusted to a concentration of 1 × 10⁷/ml using cell culture medium containing 10% fetal bovine serum, and cell viability is assessed using trypan blue (viability should be above 95%).

2.2.2 Aplastic Anemia Modeling

90 BALB/C mice, except for the 15 in the normal group, the remaining 60 mice are subjected to whole-body irradiation with ⁶⁰Co X-rays at a height of 100 cm for 1 minute at a dose of 3 Gy. Within 4 hours, 0.2 ml of a mixed cell suspension from the thymus and lymph nodes of DBA/2 mice is infused intravenously via the tail vein, with a cell count of 1 × 10⁷ cells per mouse.