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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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Histological sections from biopsies taken during laparoscopy.A – normal liver (×100, H&E), B – TAA induced fibrosis (×100, H&E), C - MCD Diet induced NASH (×40, H&E), D - TAA induced fibrosis (×40, Serius Red).
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pone-0004776-g005: Histological sections from biopsies taken during laparoscopy.A – normal liver (×100, H&E), B – TAA induced fibrosis (×100, H&E), C - MCD Diet induced NASH (×40, H&E), D - TAA induced fibrosis (×40, Serius Red).

Mentions: Liver biopsies were taken by a miniaturized biopsy forceps that was inserted through the endoscope working channel. Biopsy taking necessitated the presence of two examiners, one guiding the endoscope to the desired liver segment, the other controlling the opening and closure of the biopsy forceps. Multiple biopsies could be undertaken, each lasting approximately 30 seconds. Mild, self-contained bleeding validated that the liver tissue was indeed biopsied (video S3). No mortality was associated with the biopsy. Biopsy size was 3–7 mm in average. Time requirements for a single laparoscopy for liver examination are around 45 seconds per mouse. A biopsy prolongs the procedure by up to one extra minute. Figure 5 depicts the histological staining of representative biopsy specimens from normal liver (figure 5A, H&E stain), MCD-induced non-alcoholic liver (figure 5B) showing an area of intense lymphocytic exudate, figure 4C showing an area of severe steatosis, (H&E stain) and a liver with TAA-induced fibrosis (figure 5D, Sirius red stain). In all biopsy specimens, full correlation was observed with corresponding histological slides of livers from the same mice (data not shown).


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)

Histological sections from biopsies taken during laparoscopy.A – normal liver (×100, H&E), B – TAA induced fibrosis (×100, H&E), C - MCD Diet induced NASH (×40, H&E), D - TAA induced fibrosis (×40, Serius Red).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0004776-g005: Histological sections from biopsies taken during laparoscopy.A – normal liver (×100, H&E), B – TAA induced fibrosis (×100, H&E), C - MCD Diet induced NASH (×40, H&E), D - TAA induced fibrosis (×40, Serius Red).
Mentions: Liver biopsies were taken by a miniaturized biopsy forceps that was inserted through the endoscope working channel. Biopsy taking necessitated the presence of two examiners, one guiding the endoscope to the desired liver segment, the other controlling the opening and closure of the biopsy forceps. Multiple biopsies could be undertaken, each lasting approximately 30 seconds. Mild, self-contained bleeding validated that the liver tissue was indeed biopsied (video S3). No mortality was associated with the biopsy. Biopsy size was 3–7 mm in average. Time requirements for a single laparoscopy for liver examination are around 45 seconds per mouse. A biopsy prolongs the procedure by up to one extra minute. Figure 5 depicts the histological staining of representative biopsy specimens from normal liver (figure 5A, H&E stain), MCD-induced non-alcoholic liver (figure 5B) showing an area of intense lymphocytic exudate, figure 4C showing an area of severe steatosis, (H&E stain) and a liver with TAA-induced fibrosis (figure 5D, Sirius red stain). In all biopsy specimens, full correlation was observed with corresponding histological slides of livers from the same mice (data not shown).

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