Animal models of transplanted tumors are tumors formed by transplanting animal or human tumors into homozygous or heterozygous animals for successive generations. Tumor transplantation in healthy animals is equivalent to living tissue culture, which allows long-term preservation of tumor species for experimental use. Transplanted tumor animal models are currently the most commonly used in vivo method for antitumor drug screening and have an important role in tumor research.
Operation method
Establishment of an animal model of transplantation tumor
Materials and Instruments
Equipment: Move The establishment of transplanted tumor animal model was divided into two categories according to the type: solid tumor and ascites type, and the specific steps were as follows: . Tumor tissue block inoculation method: select tumor source animals with good growth status 7-10 days after inoculation, after execution, disinfect the skin of the inoculation site, incise the skin, puncture out the inoculated tumor block, select tumor tissues that are well survived and not necrotic, and cut the tumor tissues into small pieces of 2 mm2 in the petri dish containing sterilized PBS placed on ice. Use a sterile cannula needle to aspirate the tumor blocks and inoculate them in the axillary subcutis of homozygous recipient animals. B. Tumor cell suspension inoculation method: aseptically remove the tumor, cut the tumor into small pieces as much as possible, grind the tumor in one direction with sterile saline in a sterile glass homogenizer, filter through a strainer, dilute the tumor cell suspension with saline (1:3-1:5), count the number of viable cells by Taipan blue staining, and then use a 1 mL syringe to inoculate 0.2 mL of tumor cell solution (containing 1×106-1×107 cells). The tumor cells were inoculated with 0.2 mL of tumor cell fluid (containing 1×106-1×107 cells) using 1 mL syringe. C. Cultured cell inoculation method: After the logarithmic growth stage tumor cells were digested and demyelinated with 0.25% trypsin, centrifuged with PBS at 1000 r/min for 10 minutes, washed twice, washed off the trypsin in the cells and the serum and other components in the medium, counted the number of viable cells by TPB staining, and then made the tumor into a certain concentration of cell suspension by PBS. Use a 1 ml syringe to inoculate 0.2 mL of tumor cell solution (containing 1×106-1×107 cells). D. In situ transplantation: Take C6 cells, Wistar rats as an example: after anesthetizing the rats with 1% sodium pentobarbital (60 mg/kg) by intraperitoneal injection, the head of the rats was fixed on the brain stereotaxic apparatus to cut off the hair on the top of the head, and then sterilized by iodine and ethanol to spread the cavity towel. The scalp was incised backward longitudinally at the intersection of the medial canthus line and the mid-sagittal plane of the head for about l cm, and the skull was separated and exposed. Under the guidance of the stereotaxic instrument and according to the Batker method, the target point of the caudate nucleus was determined 1 mm behind the coronal suture and 3 mm to the right of the midline. A small hole was drilled with a dental drill without damaging the dura mater, and a 25-μl micro-syringe was used to extract 10 μl of C6 cell suspension (1 × 106 C6 cells), and the needle was inserted slowly and straightly along the bone holes until it reached 6 mm under the dura mater, and then retreated 1 mm (5 mm from the dura mater). The injection rate was 1 μl/min for 10 minutes. After the injection, the needle was left in place for 5 minutes and slowly withdrawn. The bone holes were closed with dentifrice powder. The surgical field was rinsed with saline, and the skin was sterilized after the incision was closed with a No. 4 suture. The entire inoculation process was carried out on a laminar flow ultra-clean bench, and no anti-infective treatment was required in the postoperative period, and the animals were kept in a single cage. . Select the tumor-bearing animals with good growth condition 5-7 days after inoculation, after execution, fix them in supine position, disinfect the abdominal skin, cut open the abdominal skin, and extract ascites by stabbing into the abdominal cavity with a sterile syringe, and the extracted ascites should be a milky-white viscous liquid, and it should be discarded if it is yellow or contains a lot of erythrocytes For more product details, please visit Aladdin Scientific website.
Tumor block, sterile cannulated needle, syringe, centrifuge, etc.
Reagents:
① Sterile saline;
② Trypsin;
③ PBS;
④ Taipan blue.