Introduction
A patient-derived colon cancer Xenograft (PDX) model has been established for the first time by Noble Life Sciences by implanting cryopreserved PDX fragments into a transplant compliant mouse host. PDX models have advantages over cancer cell line xenografts, and are ideal for preclinical drug screening and precision medicine. Our PDX model can provide important insight to your preclinical study.
Materials and Methods
Mouse : Athymic nude mice (Athymic Nude-Foxn1nu) Charles River
Colon PDX fragment (CCR-001, CCR-048)
Alexa Fluor 488 anti-human CD66a/c/e antibody
Method
Mice were surgically implanted with one thawed PDX fragment (2 x 2 mm) dipped in ice cold matrigel at each flank. The wound edges were stapled using autoclip staple applicator and removed 7 days post-surgery.
Tumor growth was monitored bi-weekly by palpating tumor implantation site. Tumor volumes were measured bi-weekly using digital caliper. Once the tumor reached around 750 mm3 size, mouse was humanely euthanized and tumor was extracted, sliced into small pieces (2 x 2 mm each piece) removing necrotic tissue. Tumor pieces were viably banked by placing 5 pieces per cryovial in cryopreserving medium or directly implanted in another set of mice for second PDX passage.
Results
Colon cancer PDX model was successfully made and maintained for 2 passages (Figures 1 and 2). Improved take rate with increase in passage number (Figures 1 A and Figures 2A and B). Tumors express human tumor marker (Figure 3).
Figure 1: Colon PDX model 1st passage. Tumor volume measurements of right flank of mice shown in A and body weights of mice shown in B.
Figure 2: Colon PDX model 2nd Passage. Tumor volume measurements of left flank shown in A, on right flank shown in B and the body weights of mice are shown in C.
Figure 3: Colon cancer PDX tumor (second passage) expressing Human Colon cancer marker CD66. Tumor tissue from left and right flanks after second passage were dissected, dissociated tumor cells were stained with anti-human CD66 antibody. Flow cytometry analysis of all tumors revealed expression of CD66 protein.
Model Applications:
- Platform for Preclinical drug screening
- Determination of optimal drug doses, treatment scheme and route of administration to maximize therapeutic window
- Precision Medicine
Model Read outs:
- Palpable tumor size
- Body weights
- Flow cytometry or Immune Histochemistry of tumor tissue
- In vitro assays for biomarker signature
