Model Introduction
Leukemia is a malignant proliferative disease of hematopoietic tissues, characterized by the unrestricted proliferation of nucleated cells in the bone marrow and other hematopoietic tissues, followed by infiltration into systemic organs. Acute Myeloid Leukemia (AML) models are primarily constructed through in vivo transplantation, in vitro culture, gene editing technologies, human virus mediation, and in vitro 3D culture. Common animal model classifications include induced models, transplantation models, and genetically modified models, aimed at simulating the malignant proliferation, tissue infiltration, and related clinical pathological features of AML cells in vivo.
Research Applications
This model is mainly used to explore the pathogenesis of AML, the biological behavior of leukemia cells (such as proliferation, infiltration, and metastasis), prognostic analysis of ferroptosis-related genes, and the construction of relational networks. In preclinical research, this model is widely applied in drug screening, efficacy evaluation (e.g., drugs like Cytarabine), and the study of leukemia stem cell (LSC) characteristics.
Key Points of Experimental Design
- Experimental Animal Selection: Commonly used are SPF-grade SCID mice (7–8 weeks old, body weight 20–22g) or other immunodeficient rodents.
- Cell Preparation:
- HL-60 Cells: Use RPMI-1640 medium containing 10% fetal bovine serum (FBS) to prepare a cell suspension of $1 \times 10^7 / \text{mL}$.
- KG1a Cells: Use IMDM medium containing 10% FBS, using cells in the logarithmic growth phase or sorting the $CD34+CD38-$ cell subpopulation.
- Pre-modeling Treatment: Intraperitoneal (i.p.) injection of Cyclophosphamide (100 mg/kg) for 2 consecutive days prior to inoculation to suppress the immune function of the mice.
- Inoculation Parameters: On the 3rd day post-treatment, a single-point subcutaneous injection of HL-60 cell suspension ($1 \times 10^7$ per mouse) is performed in the right abdomen.
- Dosing Intervention:
- Routine Drugs: Starting from the 3rd week after modeling, daily administration via gavage (p.o.) for 5 weeks.
- Chemotherapy Reference: Cytarabine (Ara-C) administered via tail vein injection at a dose of $100 \text{ mg}/(\text{m}^2 \cdot \text{d})$ for 5 consecutive days.
Key Monitoring Indicators
- Tumorigenicity and Survival Evaluation: Monitoring the average time to tumor formation (reference value approx. $14.63 \pm 2.74$ days), tumorigenicity rate, and average survival time.
- Hematologic Indicators: Manual counting of peripheral blood leukocytes, erythrocytes, neutrophils, lymphocytes, and total HL-60 cell count. The reference standard for successful tumorigenicity is a peripheral white blood cell count reaching $(4.03 \pm 1.92) \times 10^9 / \text{L}$.
- Pathological Infiltration Observation: Observing tumor infiltration and metastasis in visceral organs such as the liver, spleen, and lungs, particularly the degree of hepatosplenomegaly.
- Molecular and Cellular Level Detection:
- Use of RT-qPCR and Western Blot to detect the expression of relevant genes and proteins.
- Use of Flow Cytometry (FCM) to detect the purity of sorted cells (e.g., $CD34+CD38-$).
- Evaluation of cell proliferation capacity via CCK8 assay.
