Animal models of human xenograft tumors

Summary

Allogeneic animal transplantation is often difficult due to immune rejection, which often leads to graft failure. Therefore, the selection of a suitable transplantation host is the key to the success of transplantation. Human cancer cell lines cultured in vitro were successfully transplanted into nude mice, in which the human tumors still maintained their original histological morphology, unique chromosome grouping, and original characteristics to chemoradiation therapy. According to the transplantation site, they are divided into subcutaneous transplantation tumors and in situ transplantation tumors.

Operation method

Establishment of animal models of human xenograft tumors

Materials and Instruments

Animal model:
Nude mice
Material:
Human tumor cells or tissue blocks, etc.

Move

The animal models of human xenograft tumors were divided into subcutaneous transplantation tumors and intracranial in situ transplantation according to the transplantation location, and the specific steps were as follows:

(I) Subcutaneous transplantation tumor: A

. In the ultra-clean bench, the logarithmic growth stage cells were digested with 0.25% trypsin to remove the wall.

B. Centrifuge the cells with PBS at 1000 r/min for 10 minutes and wash them twice.

C. Count the number of viable cells by Tapan Blue staining, and make a certain concentration of cell suspension with PBS.

D. Use a 1 mL syringe to inoculate 0.2 mL of tumor cell solution (containing 1×105-1×107 cells) under the skin of the right (left) side of nude mice.

E. After inoculation, the nude mice were kept in an SPF-rated environment for regular observation of their spirit, diet and defecation, and the size of the tumor nodules.

(B) Intracranial in situ transplantation of tumor: A

. Animals were anesthetized intraperitoneally with 1% sodium pentobarbital, and the heads of the nude mice were fixed with the head frame of the mouse brain stereotaxic localizer in a laminar flow ultra-clean table, and the localization was corrected only to the coordinates of the "0" point.

B. A longitudinal incision of about 1 cm was made at the intersection of the medial contiguous line and the sagittal midline of the head to separate and expose the skull.

C. The location of the drill hole was determined according to the literature, and the skull was carefully drilled with a dental drill 1 mm behind the coronal suture and 2.5 mm to the right of the sagittal line, and the depth was determined to reach the dura without puncturing the dura.

D. Fix the microsampler with 5 μl of tumor cell suspension on the crossbar of the stereotactic apparatus, and enter the brain parenchyma vertically through the drilling hole. The depth of the needle was about 3.5 mm from the skull surface, and the needle was slightly returned by 0.5~1 mm before injection, then the tumor cell suspension was injected into the caudate nucleus of the right brain of the nude mice in a slow and uniform speed, and then stayed for 2 minutes before removing the needle, and then slowly withdrew the needle.

E. After the operation, bone wax was used to close the bone holes, the incision was sutured, and the remaining cells were back-cultured to detect cell viability and contamination.

F. After inoculation, all the animals were kept in SPF-grade environment, and the observation was terminated 14-20 days after inoculation or when the nude mice showed obvious neurological symptoms such as movement disorders, bowed back, agitation, hemiparesis, and so on. The observation was terminated when the animals showed obvious neurological symptoms such as movement disorders, bowed back, agitation, hemiplegia, etc.

Caveat

Nude mice are expensive to keep and require high experimental conditions. Cages, drinking water, and feed should be autoclaved and disinfected, and all experimental processes should be SPF-grade.

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