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. 2022 Sep 3;22(1):142.
doi: 10.1186/s12894-022-01093-6.

Establishment of an optimized orthotopic bladder cancer model in mice

Jinming Cai #  1 , Zhiwen Xie #  1 , Yilin Yan  1 , Zhengnan Huang  1 , Pengfei Tang  2 , Xiangqian Cao  1 , Zeyi Wang  2 , Chenkai Yang  1 , Mingyue Tan  3 , Fang Zhang  4 , Bing Shen  5
Affiliations

Establishment of an optimized orthotopic bladder cancer model in mice

Jinming Cai et al. BMC Urol. .

Abstract

Background: Bladder cancer (BC) is one of the most common malignancies of the genitourinary system. Animal models offer an important tool to explore tumour initiation, progression, and therapeutic mechanisms. Our aim is to construct an optimized orthotopic BC model which is predictable, reproducible, and convenient.

Methods: The optimized orthotopic BC model was constructed in male C57BL/6 mice utilizing microsyringes to inoculate them with a murine BC cell line (MB49). Anesthetised mice were inoculated with an MB49 cell suspension (10 µL) at approximately 5 × 106/mL. The whole process of modelling was observed and monitored every 3 days for 21 days utilizing HE staining and transabdominal ultrasonography (TUS).

Results: In this study, the model showed excellent success rates for tumour formation (96.67%) and metastatic rate (89.66%). Compared to the control group (sham operation), mice in the modelling group had serous cachexia, visible haematuresis and weight loss (all P < 0.05). The lungs, liver, ureter and kidneys were found to have tumour metastasis. Moreover, the average survival time (19.73 ± 1.69 d) of modelling mice was significantly shorter than that of the control mice (P < 0.05), which remained alive.

Conclusion: Our study established a method using microsyringes to inject murine BC cells into the bladder wall, creating a stable transplantable BC model in mice.

Keywords: Bladder cancer; MB49; Microsyringes; Orthotopic murine model.

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Conflict of interest statement

All authors declared that they have no conflict of interest.

Figures

Fig. 1
Fig. 1
Microsyringes in scattered (a) and assembled (b)
Fig. 2
Fig. 2
The operation procedures of orthotopic transplantation. a The lower abdomen area was depilated and disinfected. b The bladder was dissected and identify vascular-enriched area. c Gentle inject cells. d Observation after injection. e Close the lower abdomen and disinfect
Fig. 3
Fig. 3
Exhibition of murine hematuresis (a) and abdominal organs (b)
Fig. 4
Fig. 4
TUS of a mouse bladder tumor and liver. a The growth of bladder tumor can be observed and measured longitudinally via TUS. b The liver can be monitored by TUS during the observation of bladder tumor
Fig. 5
Fig. 5
Survival time of the modelling and the sham operation
Fig. 6
Fig. 6
Comparison of average bladder wet weight (a) and average mouse weight (b) and metastasis analysis (c, d)
Fig. 7
Fig. 7
Pathological images and TUS of mice bladder and liver in the control and modelling group. a, b Pathological images (HE staining) of bladder tumor in the control group (sham operation) and the modelling group. c, d Bladder and liver of TUS of bladder tumor in the control group (sham operation) and the modelling group. e Pathological images (HE staining) of liver in the control group
Fig. 8
Fig. 8
Pathological images of spleen (a), lung (b), liver (c), heart (d), bladder (e), ureter (f), kidney (g)
See this image and copyright information in PMC

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