Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Aug 4:38:e382823.
doi: 10.1590/acb382823. eCollection 2023.

Pancreatic cancer orthotopic graft in a murine model

Milena Muzzolini  1   2 , Ismahane Belhabib  3 , Victoire Cardot  4 , Annemilaï Tijeras-Raballand  5 , Cindy Neuzillet  1   6   7 , Corinne Bousquet  3 , Renato Micelli Lupinacci  1   6 , Christine Jean  3
Affiliations

Pancreatic cancer orthotopic graft in a murine model

Milena Muzzolini et al. Acta Cir Bras. .

Abstract

Purpose: Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers with increasing incidence. Even if progress have been made, the five-year overall survival remains lower than 10%. There is a desperate need in therapeutic improvements. In the last two decades, new in-vitro models have been developed and improved, including tridimensional-culture spheroids and organoids. However, animal studies remain mandatory in the upscaling before clinical studies. Orthotopic and syngeneic grafting is a robust model to test a drug efficiency in a tumor and its microenvironment.

Methods: We described a method for orthotopic and syngeneic graft of KRAS mutated, p53 wildtype, 8305 cells in a C57BL/6J mouse model.

Results: With this microsurgical method, 30 mice were grafted, 24 by a junior and six by a senior, resulting in 95,8 and 100% of (partial and total) successful tumoral implantation, respectively. Twenty mice underwent ultrasound follow-up. It was an efficient method for the tumoral growth evaluation. At day 16 after grafting, 85% of the tumors were detectable by ultrasound, and at day 22 all tumors were detected.

Conclusions: The presented method appears to be a robust and reliable method for pre-clinical studies. A junior master student can provide positive results using this technique, which can be improved with training.

PubMed Disclaimer

Conflict of interest statement

Conflict of interest: Nothing to declare.

Figures

Figure 1
Figure 1. Orthotopic graft for pancreatic cells. (a) Mouse installation, black arrow shows the facial mask. (b) Pointer for a good incision are the grill cost and the beginning of the tail. (c) Incision 1 cm away from the grill cost. (d) Incision between two clinches. (e) Subcutaneous detachment. (f) Peritoneal incision. (g) Exposition and direction of the needle for injection. Red marker represents the splenic artery, above is the spleen and beneath is the pancreas. (h) Verification of the bubble formation (blue arrow). (i) Peritoneum closure. (j) Skin closure with a stapler.
Figure 2
Figure 2. Schematic representation of the planned experiments of grafted mice and their follow-up.
Figure 3
Figure 3. Mice follow up. (a) Mean weight and standard deviation based on initial weight (day 0): Sac 500 mm3 (blue): mice sacrificed when tumor volume reached 500 mm3, Sac 31 days (green): mice sacrificed at day 31, and Sac 200 mm3 (orange): mice sacrificed when tumor volume reached 200 mm3. (b) Mean tumoral volume and replicates estimated using ultrasound for each group, on the cohort grafted by a junior operator, Sac 500 mm3 (blue): mice sacrificed when tumor volume reached 500 mm3, Sac 31 days (green): mice sacrificed at day 31 and Sac 200 mm3 (orange): mice sacrificed when tumor volume reached 200 mm3. (c) Mean tumoral volume and replicates estimated using ultrasound on the cohort grafted by a senior team. (d) Correlation between tumor volume estimation using ultrasound and tumor weight and standard linear regression (y = 1,3403 x, R2 = 0,8529).
See this image and copyright information in PMC

References

    1. Rahib L, Smith BD, Aizenberg R, Rosenzweig AB, Fleshman JM, Matrisian LM. Projecting Cancer Incidence and Deaths to 2030: The Unexpected Burden of Thyroid, Liver, and Pancreas Cancers in the United Statesconroy. Cancer Res. 2014;74(11):2913–2921. doi: 10.1158/0008-5472.CAN-14-0155. - DOI - PubMed
    1. Vincent A, Herman J, Schulick R, Goggins M. Pancreatic cancer. Lancet Lond Engl. 2011;378(9791):607–620. doi: 10.1016/S0140-6736(10)62307-0. - DOI - PMC - PubMed
    1. Neuzillet C, Gaujoux S, Williet N, Bachet JB, Bauguion L, Colson Durand L, Conroy T, Dahan L, Gilabert M, Huguet F, Marthey L, Meilleroux J, De Mestier L, Napoleon B, Portales F, Sa Cunha A, Schwartz L, Taieb J, Chibaudel B, Bouche O, Hammel P. Pancreatic cancer: French clinical practice guidelines for diagnosis, treatment and follow-up (SNFGE, FFCD, GERCOR, UNICANCER, SFCD, SFED, SFRO, ACHBT, AFC) Dig Liver Dis Off J Ital Soc Gastroenterol Ital Assoc Study Liver. 2018;50(12):1257–1271. doi: 10.1016/j.dld.2018.08.008. - DOI - PubMed
    1. Ducreux M, Cuhna AS, Caramella C, Hollebecque A, Burtin P, Goere D, Seufferlein T, Haustermans K, Van Laethem JL, Conroy T, Arnold D. Cancer of the pancreas: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol Off J Eur Soc Med Oncol. 2015;26(Suppl. 5):V56–V68. doi: 10.1093/annonc/mdv295. - DOI - PubMed
    1. Neuzillet C, Gaujoux S, Williet N, Bachet JB, Bauguion L, Colson L Durand, Conroy T, Dahan L, Gilabert M, Huguet F, Marthey L, Meilleroux J, De Mestier L, Napoleon B, Portales F, Sa Cunha A, Schwartz L, Taieb J, Chibaudel B, Bouche O, Hammel P. Pancreatic cancer: French clinical practice guidelines for diagnosis, treatment and follow-up (SNFGE, FFCD, GERCOR, UNICANCER, SFCD, SFED, SFRO, ACHBT, AFC) Dig Liver Dis Off J Ital Soc Gastroenterol Ital Assoc Study Liver. 2018;50(12):1257–1271. doi: 10.1016/j.dld.2018.08.008. - DOI - PubMed