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Utilization of murine laparoscopy for continuous in-vivo assessment of the liver in multiple disease models.

Shapira Y, Katz M, Ali M, Kaplan M, Brazowski E, Halpern Z, Elinav E - PLoS ONE (2009)

Bottom Line: Current strategies for follow up of murine models of liver disease are flawed by inability to continuously monitor disease progression in the tissue level, and necessitate sacrifice of animals for tissue sampling.In this study we aimed at developing a safe repetitive tool for sampling livers in vivo, by utilization of a miniaturized endoscopy system for laparoscopic liver biopsies and for injection of tumor cells into livers.The system enables safe and repeated liver biopsies in mice and rats, yielding adequate tissue for histological staining and RNA extraction.

View Article: PubMed Central - PubMed

Affiliation: Institute for Gastroenterology and Liver Disease, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel.

ABSTRACT

Background: Current strategies for follow up of murine models of liver disease are flawed by inability to continuously monitor disease progression in the tissue level, and necessitate sacrifice of animals for tissue sampling.

Aims: In this study we aimed at developing a safe repetitive tool for sampling livers in vivo, by utilization of a miniaturized endoscopy system for laparoscopic liver biopsies and for injection of tumor cells into livers.

Results: We report the development of a protocol for murine laparoscopy that allows repeated visualization of murine intra-abdominal organs. The system enables safe and repeated liver biopsies in mice and rats, yielding adequate tissue for histological staining and RNA extraction. In addition, injection of tumor cells into livers facilitates under-vision implantation of hepatic tumors in liver, followed by visualization of tumor growth.

Conclusions: Murine laparoscopy may be employed as a novel imaging modality for continuous assessment and manipulation of chronic liver disease models.

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Related in: MedlinePlus

Liver tumors implanted during murine laparoscopy.A. In-vivo imaging (IVIS 100) of hepatic luciferase signal in sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. B. In-vivo imaging (ZIESS Axioimager-M) of hepatic GFP fluorescence in sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. C. Repeated GFP imaging (ZIESS Axioimager-M) in excised livers of sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. D. Laparoscopic appearance of liver-infiltrating GFP-HCC tumors 7 & 14 days following hepatic tumor cell implantation.
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pone-0004776-g007: Liver tumors implanted during murine laparoscopy.A. In-vivo imaging (IVIS 100) of hepatic luciferase signal in sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. B. In-vivo imaging (ZIESS Axioimager-M) of hepatic GFP fluorescence in sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. C. Repeated GFP imaging (ZIESS Axioimager-M) in excised livers of sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. D. Laparoscopic appearance of liver-infiltrating GFP-HCC tumors 7 & 14 days following hepatic tumor cell implantation.

Mentions: One of the most attractive usages of murine laparoscopy is the accurate guided implantation of tumor cells within target organs. To demonstrate such ability, 5×106 GFP or Fluorscin-positive BNL1ME hepatocellular carcinoma cells were suspended in 200 microliter of HBSS and directly injected into the right hepatic lobe through the murine laparoscope working channel (video S4). Using the IVIS in-vivo imaging system for luminescence detection, the fluorescin-positive tumor could be visualized within the liver as soon as 3 days after implantation (data not shown), and was brightly-positive in all luciferase-tumor harboring mice, but in non of the sham-operated or GFP-positive mice, 10 days after tumor implantation (figure 7A). Inversely, using the Carl Zeiss Axioimager-M fluorescent microscopy system for in-vivo detection of GFP signals, in the two mice in which a GFP-positive tumor was implanted, intense fluorescent signal could be detected, while no signal was omitted from either sham or fluorescin-positive tumors (figure 7B). Liver excision from all mice confirmed that the GFP signal, detected in-vivo, indeed originated from the hepatic tumors (figure 7C). Tumors could be repetitively examined and biopsied using routine weekly laparoscopy (figure 7d). In our hands, tumor implantation by left lateral laparoscopy followed by repeated visualization of tumor growth by right lateral thoracotomy yield the most reproducible results.


Utilization of murine laparoscopy for continuous in-vivo assessment of the liver in multiple disease models.

Shapira Y, Katz M, Ali M, Kaplan M, Brazowski E, Halpern Z, Elinav E - PLoS ONE (2009)

Liver tumors implanted during murine laparoscopy.A. In-vivo imaging (IVIS 100) of hepatic luciferase signal in sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. B. In-vivo imaging (ZIESS Axioimager-M) of hepatic GFP fluorescence in sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. C. Repeated GFP imaging (ZIESS Axioimager-M) in excised livers of sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. D. Laparoscopic appearance of liver-infiltrating GFP-HCC tumors 7 & 14 days following hepatic tumor cell implantation.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2651645&req=5

pone-0004776-g007: Liver tumors implanted during murine laparoscopy.A. In-vivo imaging (IVIS 100) of hepatic luciferase signal in sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. B. In-vivo imaging (ZIESS Axioimager-M) of hepatic GFP fluorescence in sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. C. Repeated GFP imaging (ZIESS Axioimager-M) in excised livers of sham operated, GFP-HCC implanted or luciferase-HCC implanted mice. D. Laparoscopic appearance of liver-infiltrating GFP-HCC tumors 7 & 14 days following hepatic tumor cell implantation.
Mentions: One of the most attractive usages of murine laparoscopy is the accurate guided implantation of tumor cells within target organs. To demonstrate such ability, 5×106 GFP or Fluorscin-positive BNL1ME hepatocellular carcinoma cells were suspended in 200 microliter of HBSS and directly injected into the right hepatic lobe through the murine laparoscope working channel (video S4). Using the IVIS in-vivo imaging system for luminescence detection, the fluorescin-positive tumor could be visualized within the liver as soon as 3 days after implantation (data not shown), and was brightly-positive in all luciferase-tumor harboring mice, but in non of the sham-operated or GFP-positive mice, 10 days after tumor implantation (figure 7A). Inversely, using the Carl Zeiss Axioimager-M fluorescent microscopy system for in-vivo detection of GFP signals, in the two mice in which a GFP-positive tumor was implanted, intense fluorescent signal could be detected, while no signal was omitted from either sham or fluorescin-positive tumors (figure 7B). Liver excision from all mice confirmed that the GFP signal, detected in-vivo, indeed originated from the hepatic tumors (figure 7C). Tumors could be repetitively examined and biopsied using routine weekly laparoscopy (figure 7d). In our hands, tumor implantation by left lateral laparoscopy followed by repeated visualization of tumor growth by right lateral thoracotomy yield the most reproducible results.

Bottom Line: Current strategies for follow up of murine models of liver disease are flawed by inability to continuously monitor disease progression in the tissue level, and necessitate sacrifice of animals for tissue sampling.In this study we aimed at developing a safe repetitive tool for sampling livers in vivo, by utilization of a miniaturized endoscopy system for laparoscopic liver biopsies and for injection of tumor cells into livers.The system enables safe and repeated liver biopsies in mice and rats, yielding adequate tissue for histological staining and RNA extraction.

View Article: PubMed Central - PubMed

Affiliation: Institute for Gastroenterology and Liver Disease, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel.

ABSTRACT

Background: Current strategies for follow up of murine models of liver disease are flawed by inability to continuously monitor disease progression in the tissue level, and necessitate sacrifice of animals for tissue sampling.

Aims: In this study we aimed at developing a safe repetitive tool for sampling livers in vivo, by utilization of a miniaturized endoscopy system for laparoscopic liver biopsies and for injection of tumor cells into livers.

Results: We report the development of a protocol for murine laparoscopy that allows repeated visualization of murine intra-abdominal organs. The system enables safe and repeated liver biopsies in mice and rats, yielding adequate tissue for histological staining and RNA extraction. In addition, injection of tumor cells into livers facilitates under-vision implantation of hepatic tumors in liver, followed by visualization of tumor growth.

Conclusions: Murine laparoscopy may be employed as a novel imaging modality for continuous assessment and manipulation of chronic liver disease models.

Show MeSH
Related in: MedlinePlus