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Abstract
Objectives of this study were to establish a leukemia mouse model in the Balb/c mouse based upon the A20 cell line (murine B-lymphoma/leukemia cell line, H-2d). Here we demonstrate for the first time that A20 cells were infiltrated into tissue and bone marrow, thereby evaluate the feasibility of using A20 leukemic cells as a leukemia model. In the study, changes of behavior, survival rate and histological changes of major organs after intravenous injection of A20 cells (1×105, 1×106 or 1×107) into Balb/c mice were observed. After inoculation of 1×106 cells, animals survived up to 38.3 days, although there were no significant correlation between the number of injected cells and life-span. At 21 and 28 days postinjection, both hematoxylin-eosin and CD45R immunohistochemical stains showed diffuse large B-cell lymphoma in the liver. FACS analysis was performed after injection of fluorescent nanomaterial (MNPs@SiO2 RITC)-labeled A20 cells. The labeled A20 cells were detected in bone marrow from 6 hours post-inoculation, indicative of the cellular infiltration. This is the first study that demonstrated the invasion of A20 cells into the bone marrow of Balb/c model using A20 cells. With the occurrence of systemic lesions following metastasis of the cells into lymph nodes and neighboring tissues via bone marrow infiltration, it is suggested that the A20 cell-inoculated Balb/c miouse could be an animal model of acute lymphocytic leukemia.
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Figure 1.
Morphology and CD45R expression of A20 cell line, showing features immediately (A) and 48 hours after seeding (B) (scale bar: 50 µm), Wright's staining (C) (scale bar: 100 µm), and CD45R expression on the cell surface (D).
Figure 2.
Survival rate of A20 cell-transplanted Balb/c mice. Three different numbers (1×105, 1×106 or 1×107 cells) of A20 leukemia cells were intravenously injected into Balb/c mice. Each group consisted of 6–13 mice.
Figure 3.
Comparison of the livers of control and A20 cell-inoculated mice. Livers were removed at 28 days post-injection. The surface of control liver is smooth and soft, while distinct nodules are visible on the A20 cell-transplanted mice (A-D). Histological examination showed diffuse or nodular infiltration of lymphoma cells in the liver parenchyma of A20 cell-transplanted mice. The infiltrated A20 cells were confirmed by H&E (E-G) and immunohistochemical staining on CD45R (H-K). Scale bar: 500 µm.
Figure 4.
Detection of fluorescent nanoparticle (MNPs@SiO2 RITC)-labeled A20 cells in vitro and in vivo. RITC signals in the labeled cells were detected by fluorescence microscopy (A) and FACS (B). In vivo, the infiltrations of A20 cells in PBMC (C) and bone marrow (D) were analysis by FACS. Data at every time point in each group consisted of 3 mice.
Table 1.
Body and tissue weights of A20 cell-transplanted Balb/c mice
| Treatment (cells) | Survival (days) | Body weight (g) | Liver weight (g) | Spleen weight (g) | ||
|---|---|---|---|---|---|---|
| 0 week | 3 week | 4 week | 4 week | 4 week | ||
| PBS | ∞ | 100.0±4.5 | 108.5±7.1 | 111.0±5.4 | 1.351±0.051 | 0.074±0.006 |
| 1x105 | 42.5±6.2 | 100.0±5.4 | 107.5±6.3 | 110.9±7.6 | 1.374±0.021 | 0.078±0.005 |
| 1x106 | 38.3±5.4 | 100.0±3.7 | 104.1±6.4 | 110.1±4.1 | ∗1.590±0.082∗ | 0.087±0.010 |
| 1x107 | 36.3±5.3 | 100.0±1.6 | 107.6±3.1 | 103.4±4.7 | ∗1.985±0.167∗ | ∗0.134±0.121∗ |
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