Search

Murine xenograft model   

Download PDF How to cite Favorites 1 Q&A Share your feedback Cited by

In this protocol

Abstract

This experiment is used to test the efficacies of chemo treatments or gene therapy in an in vivo system. In this protocol, the mouse xenograft model is used.

Materials and Reagents

  1. Severe combined immunodeficiency (SCID) mice
  2. NaCl
  3. Pluronic acid
  4. Phosphate buffered saline (PBS)
  5. HBSS solution

Equipment

  1. 1 ml syringe with 22-24 gauge of needls
  2. Bioluminescent imaging instrument (in university core facility)

Procedure

  1. For single tumor cell (or any kind of tumor cells)
    1. Six-week-old female in-bred Fox Chase SCID mice were obtained from Charles River Laboratories (Hartford, CT, USA). Animals were handled according to a protocol approved by the Institutional Animal Care and Use Committee at our university.
    2. Mice were allowed to acclimate to animal housing, and xenografts were developed by subcutaneously injecting 5 x 106 cancer cells in murine flank bilaterally. Twice weekly, tumor volume was determined using digital caliper measurements and the formula:
      large diameter2 x small diameter
                         2
    3. After 14 days, all mice had measurable tumours and were sorted into treatment and control groups with equal number of animals (n=5). Treatment group mice received some dose of chemodrugs dissolved in vehicle (0.1 M NaCl, 0.05% pluronic acid in PBS) per treatment while control mice received vehicle only.
    4. All mice received 10 intraperitoneal injections over a 14-day period (cycle: Five treatment days followed by two non-treatment days). After 14 days (2 cycles), mice were killed. So the whole experiment lasts 28 days.
    5. The tumor sample can be used either for extract RNA, protein or immunohistochemistry staining.

  2. For interaction of tumor and Stella cells (for cancers containing big volume of stella cells)
    --using pancreatic cancer model.
    1. Bioluminescent imaging was done weekly to follow the luciferase signal from BxPC3 cells.
    2. Mice were sacrificed and tumors were harvested and measured.
      1. BxPC3-FL alone, either 0.5 x 106 or 1 x 106 per mouse.
      2. BxPC3-FL with immortalized HPSCs (human pancreatic stellate cells), in varying tumor-to-stroma ratios (1:1, 1:1, or 1:5).
      3. Immortalized HPSCs alone (0.5 x 106 or 1 x 106). All cell suspensions, including the mixture of BxPC3 and HPSCs, were injected in a 50 μl volume of HBSS.
    3. Statistical analysis was done using GraphPad Prism (GraphPad). Comparisons were made using two-tailed Student’s t test and significant difference was defined as P < 0.05. Data are shown as mean ±SE.

Acknowledgments

This protocol was adapted from Hwang et al. (2008).

References

  1. Hwang, R. F., Moore, T., Arumugam, T., Ramachandran, V., Amos, K. D., Rivera, A., Ji, B., Evans, D. B. and Logsdon, C. D. (2008). Cancer-associated stromal fibroblasts promote pancreatic tumor progression. Cancer Res 68(3): 918-926.
Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.
How to cite: Liu, F. (2012). Murine xenograft model. Bio-protocol Bio101: e221. DOI: 10.21769/BioProtoc.221.
Q&A

Please login to post your questions/comments. Your questions will be directed to the authors of the protocol. The authors will be requested to answer your questions at their earliest convenience. Once your questions are answered, you will be informed using the email address that you register with bio-protocol.
You are highly recommended to post your data including images for the troubleshooting.

You are highly recommended to post your data (images or even videos) for the troubleshooting. For uploading videos, you may need a Google account because Bio-protocol uses YouTube to host videos.

Q1. Which part in mouse body do you expect to see the tumors? Does it depend on types of injected tumor cells or injection location?

Q2. Can you elaborate in more details how you visualize the tumors in step 1-(2)?
7/8/2012 12:30:51 PM Reply
FengZhi Liu
School of Biomedical Sciences, Thomas Jefferson University, USA

The mouse usually is SCID mouse. Any tumor can grow if you use enough amount. The side is subcutaneous on the back of neck. You just hod the skin with thumb and index and inject the tumor cells.

The tumors take some time to grow to visible. Some tumors grow fast, and may takes one week, some not, it depends. you just measure the sizes in different groups with a gauze.

7/8/2012 9:24:55 PM