Patent Description:
Conditional reprogramming (CR) is a cell culture technique that can be used to rapidly and efficiently establish patient-derived cell cultures from both normal and diseased cells, including tumor cells. The greatest advantage of CR is its rapid and efficient expansion of cell cultures from patient tissue samples. This allows researchers to screen tumors for sensitivity to anticancer drugs or immunotherapies quickly enough to provide the information for clinical use.

The article "<NPL>. ) concerns re-progammed tumor prostate epithelial cells used as a cell model for studying human prostate cancer. More particular, it discloses a method involving the step of cultivating said cells in F-medium consisting of <NUM>% Ham's F-<NUM> nutrient mix and <NUM>% complete DMEM, supplemented with 25ng/ml hydrocortisone, <NUM>/ml insulin, <NUM> nmol/ml cholera toxin, <NUM> ng/ml Fungizone, <NUM> ng/ml epidermal growth factor, <NUM>/ml gentamicin and <NUM>µmol/L ROCK inhibitor Y-<NUM>.

The article "<NPL>) demonstrate a method comprising the step of cultivating the cells in the presence of the ROCK inhibitor Y-<NUM> in order to produce conditionally reprogrammed primary tumor cells.

The patent <CIT> discloses an improved method for the preparation of reprogrammed cancer cells. The method therein comprises culturing the cells in a medium comprising hydrocortisone, cholera toxin, Insulin and on growth arrested feeder cells.

Testing an anti-tumor drug in an animal model for its efficacy is important as such animal model is important for anti-tumor drug screening. Such an animal model will mimic the in vivo environment of patients, reflect the patient's response and thus is more effective. To establish an animal models a person skilled in the art usually need to obtain sufficiently large number of tumor cells. However, tumor samples obtained from patients may contain very small or even trace amount of tumor cells depending on how they are obtained. For example, a tumor sample obtained from needle biopsy may contain very small amount of tumor cells and thus very difficult for a person skilled in the art to use them to establish a desired animal model for screening anti-tumor drugs.

Tumor cell immortalization through conditional reprogrammmg <NUM> an invaluable tool to generate propagating tumor cells for cell-based diagnostics, drug sensitivity assay and bio-banking in vitro. However, how to effectively and successfully cultivate primary tumor cells obtained from patients, especially low or even trace amount of cell samples from e.g. a needle biopsy, is still a challenge.

To solve the above problem, the inventor has successfully conditionally reprogrammed primary tumor cells from a number of tumor types towards immortalization and use the conditionally reprogrammed tumor cells to establish an animal model for testing anti-tumor drugs. The primary tumor cells are obtained from surgery tissues, biopsy or needle biopsy samples. The conditionally reprogrammed primary tumor cells obtained in the present invention exhibited typical colonized growth, which is well maintained upon cryo-frozen. In some cases, the cells can be passaged for multiple times and become useful cell lines. Like primary tumor cells, conditional reprogrammed tumor cells are reliable to test drug sensitivity in vitro. The inventor has successfully implanted conditional reprogrammed tumor cells (CRC) into an animal model for testing drug efficacy, and screened anti-tumor drugs using the animal model. Also described herein is the use of conditional reprogrammed tumor cells in drug efficacy tests.

One aspect of the present invention provides a method for obtaining a mouse model for screening anti-tumor drugs, comprising.

In embodiments, the tumor biopsy sample is a needle biopsy sample. In embodiments, the concentration of the primary tumor cells in the sample obtained from a needle biopsy is less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM>,<NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL or less, such as less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL. In embodiments, the primary tumor cell is cultivated with a feeder cell in the composition of step (<NUM>), in particular the feeder cell is a mouse embryonic fibroblast (MEF) cell. In embodiments, the MEF cells have been treated with mitomycin C.

The animal in the present invention is a mouse, in particular an immunodeficient mouse, such as a nude mouse. In embodiments, the primary tumor cell obtained in step (<NUM>) was implanted in to the animal in a hollow fiber. In embodiments, the hollow fiber is made of modified polyvinylidene fluoride, more particularly the hollow fiber is made of modified polyvinylidene fluoride and has a cut-off value of <NUM>,<NUM> Dalton.

One aspect of the present invention provides a method for obtaining a mouse model for screening anti-tumor drugs, comprising (<NUM>) cultivating a primary tumor cell obtained from a tumor biopsy sample of a patient in a composition comprising:.

(<NUM>) implanting the primary tumor cell obtained from step (<NUM>) into an animal, wherein the animal is a mouse. In embodiments, the tumor biopsy sample is a needle biopsy sample. In embodiments, the concentration of tumor cells in the sample obtained from the needle biopsy is less than about <NUM>*<NUM><NUM>/mL.

The concentration of hydrocortisone in the composition of step (<NUM>) can be <NUM>-<NUM>/L, in particular about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L.

The concentration of insulin in the composition of step (<NUM>) can be <NUM>-<NUM>/L, in particular about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L.

The concentration of cholera toxin in the composition of step (<NUM>) can be <NUM>-<NUM>µg/L, in particular about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L.

The concentration of adenine in the composition of step (<NUM>) can be <NUM>-<NUM>/L, in particular about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>-<NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L, about <NUM>/L.

The concentration of EGF in the composition of step (<NUM>) can be <NUM>-<NUM>µg/L, in particular about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>-<NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L, about <NUM>µg/L.

The concentration of Y-<NUM> in the composition of step (<NUM>) can be <NUM>-<NUM>µmol/L, in particular about <NUM>-20µmol/L, about <NUM>-18µmol/L, about <NUM>-16µmol/L, about <NUM>-12µmol/L, about <NUM>-llµmol/L, about <NUM>-<NUM>. 5µmol/L, about 2µmol/L, about 4µmol/L, about 6µmol/L, about 7µmol/L, about 8µmol/L, about 9µmol/L, about lOµmol/L, about llµmol/L, about 12µmol/L, about 14µmol/L, about 16µmol/L, about 18µmol/L, about 20µmol/L, about 25µmol/L, about 30µmol/L.

The concentration of FBS the composition of step (<NUM>) can be <NUM>-<NUM>%, in particular about <NUM>-<NUM>%, about <NUM>-<NUM>%, about <NUM>-<NUM>%, about <NUM>-<NUM>%, about <NUM>-<NUM>%, about <NUM>-<NUM>%, about <NUM>-<NUM>%, about <NUM>%, about <NUM> %, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM>%, about <NUM> %, about <NUM>%, about <NUM>%.

In embodiments, the present invention provides a method for obtaining an animal model for screening anti-tumor drugs, comprising (<NUM>) cultivating a primary tumor cell obtained from a tumor biopsy sample of a patient in a composition comprising:.

(<NUM>) implanting the primary tumor cell obtained from step (<NUM>) into an animal, wherein the animal is a mouse. In embodiments, the tumor biopsy sample is a needle biopsy sample. In embodiments, the concentration of the primary tumor cells in the sample obtained from a needle biopsy is less than about <NUM>*<NUM><NUM>/mL,
less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM>*<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM>,<NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL or less, such as less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL, less than about <NUM> *<NUM><NUM>/mL. In embodiments, the primary tumor cell is cultivated with a feeder cell in the composition of step (<NUM>), in particular the feeder cell is a mouse embryonic fibroblast (MEF) cell. In embodiments, the MEF cells have been treated with mitomycin C.

In embodiments, the animal in the method of the present invention is a mouse, in particular an immunodeficient mouse, such as a nude mouse. In embodiments, the primary tumor cell obtained in step (<NUM>) was implanted in to the animal in a hollow fiber. In embodiments, the hollow fiber is made of modified polyvinylidene fluoride, more particularly the hollow fiber is made of modified polyvinylidene fluoride and has a cut-off value of <NUM>,<NUM> Dalton.

The tumor cells can be obtained from a variety of tumor types, including but not limited to tumors in the digestive tract (such as the stomach, intestines, duodenum, colon, pancreas, etc.), the breast, lung, liver, and endocrine glands (such as the adrenal gland, parathyroid gland, pituitary, testis, ovaries, and thymus, thyroid gland), urinary and reproductive systems (such as kidney, bladder, ovary, testis, prostate, etc.), skeletal muscle system (such as bone, smooth muscle, striated muscle, etc.), skin, and so on. For example, the tumor cells can be derived from gastric cancer, biopsy specimen of colorectal cancer and lung cancer.

The conditionally reprogrammed primary tumor cells can be implanted into the animal via a syringe or other methods or devices known in the art. In embodiments, the conditionally reprogrammed primary tumor cells are implanted into an animal by a method for producing a PDX model for tumor growth in vivo. See e.g. "<NPL>, " <NPL>, Maria C. Villarroel. In embodiments, the conditionally reprogrammed primary tumor cells were transferred into a hollow-fiber tube, which is then implanted into an animal. The hollow fiber can be made of modified polyvinylidene fluoride and has a cut-off value of <NUM>,<NUM> Dalton. In the present invention, a mini-PDX device in the refers to a hollow fiber can be made of modified polyvinylidene fluoride and has a cut-off value of <NUM>,<NUM> Dalton, which may contain the primary tumor cells obtained from a patient and be implanted into a candidate animal, e.g. implanted into a candidate animal subcutaneously. In the present invention, a mini-PDX model refers to an animal that has been implanted with the mini-PDX device of the present invention. The mini-PDX animal model can be used in the methods/tests described in the present invention.

The present invention will be better described below with reference to the accompanymg drawings.

Medium contains the following ingredients:.

The medium was then properly kept in a cool place for use.

MEF (mouse embryonic fibroblast) cells were isolated from e13. <NUM> embryos of C57 mouse and were grown in DMEM supplemented with <NUM>% FBS. Isolated MEF cells in <NUM>-<NUM> passages were treated with mitomycin C (I0µg/ml) for <NUM> and were washed with PBS. The treated MEF were harvested and cryopreserved as feeder cell.

Conditional reprograming cells from MDX079, MDX083, MDX095, MDX107, MDX123 were collected separately. MDX079 originated from a female lung cancer patient, MDX083 model originated from a <NUM>-year old male adenoid cystic carcinoma patient, MDX095 model originated from a <NUM>-year old male peritoneal malignant tnesoshelioma patient, MDX107 model originated from a <NUM>-year old male glioblastoma patient, MDX123 model originated from a <NUM>-year old male colonrectal cancer patient.

Reverse microscope DMIL, LEICA. Balance ALC-<NUM>, Acclulab. Microbalance BSA224S, Sartorius. Centrifuge 5810R, Eppendorf.

Mini-PDX device: a hollow fiber made of modified polyvinylidene fluoride which has a cut-off value of <NUM>,<NUM> Dalton, internal diameter <NUM>-<NUM>. Cut into desired length.

The cells suspension was filled into Mini-PDX device and the devices were inoculated subcutaneously into both flanks of Nu/Nu-nude mice, to establish the Mini-PDX model. The inoculation day was defined as day <NUM>. Mice were randomized in groups according to the bodyweights and the treatments were initiated at day <NUM>. The test article administration and the mini-PDX device numbers in each study group are shown in the following experimental design tables.

The protocol and any amendment(s) or procedures involving the care and use of animals in this study were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of Shanghai LIDE prior to conduct. During the study, the care and use of animals were conducted in accordance with the regulations of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). After inoculation, the animals were checked daily for morbidity and mortality. At the time of routine monitoring, the animals were checked for any effects of tumor growth and treatments on normal behavior such as mobility, food and water consumption, body weight gain/loss (body weight were measured twice weekly or every other day), eye/hair matting and any other abnormal effect. Death and observed clinical signs were recorded on the basis of the numbers of animals within each subset.

In MDX079 Mini-PDX models, Docetaxel and Carboplatin combination treatment (T/C%=-<NUM> %) could result in a significant decrease in tumor cell viability; Gemcitabine and Carboplatin combination treatment (T/C%=<NUM>%) also shown an anti-tumor activity; Pemetrexed and Carboplatin combination treatment (T/C%=<NUM>%) treatment, Pemetrexed, Carboplatin and Bevazulimab combination treatment (T/C%=<NUM>%) treatment, Etoposide and Carboplatin combination treatment (T/C%=<NUM>%) shown no anti-tumor activity. In MDX083 Mini-PDX models, all the treatment group include Paclitaxel and Cisplatin combination treatment (T/C%=<NUM>%), Gemcitabine and Cisplatin combination treatment (T/C%=<NUM> %), Docetaxel and Carboplantin combination treatment (T/C%=<NUM>%), <NUM>-Fu and Cisplatin combination treatment (T/C%=<NUM>%), Epirubicin, <NUM>-Fu and Cisplatin combination treatment (TIC%=<NUM>%) could result in a decrease in tumor cell viability.

In MDX095 Mini-PDX models, <NUM>-Fu treatment (T/C%=<NUM>%), Paclitaxel treatment (T/C%=<NUM>%) result in a little decrease in tumor cell viability; Gemcitabine and Cisplatin combination treatment (T/C%=<NUM>%) shown no anti-tumor activity, Cisplatin treatment (T/C%=<NUM>%), Epirubicin treatment (T/C%=<NUM>%), Etoposide treatment (T/C%=<NUM>%) shown no anti-tumor activity.

In MDX107 Mini-PDX models, Temozolomide treatment shown no anti-tumor activity. In MDX123 Mini-PDX models, <NUM>-Fu treatment (T/C%=<NUM>%), Oxaliplatin treatment (T/C%=<NUM> %), Irinotecan treatment (T/C%=<NUM>%), Bevazulimab treatment (T/C%=<NUM>%) could result in a decrease in tumor cell viability; Raltitrexed treatment (T/C%=<NUM>%), Bevazulimab treatment (T/C%=<NUM> %) shown no anti-tumor activity.

Conditional reprograming cells from MDX133, MDX154, MDX164, MDX165, MDX168, MDX169, MDX174, MDX186, MDX189, MDX203 were collected separately. MDX133 originated from a <NUM>-year old male gastric cancer patient, MDX154 model originated from a female gallbladder carcinoma patient, MDX164 model originated from a <NUM>-year old male glioblastoma patient, MDX165 model originated from a <NUM>-year old male glioblastoma patient, MDX168 model originated from a <NUM>-year old female gallbladder carcinoma patient, MDX169 model originated from a <NUM>-year old male lung cancer patient, MDXI74 model originated from a <NUM>-year old male lung cancer patient, MDX186 model originated from a <NUM>-year old female pancreatic cancer patient, MDX189 model originated from a <NUM>-year old male esophagus cancer patient, MDX203 model originated from a <NUM>-year old male gallbladder carcinoma patient.

Cages: Made of polycarbonate. The size is <NUM> x <NUM> x <NUM>. The bedding material is corn cob, which is changed twice per week.

Diet: Animals had free access to irradiation sterilized dry granule food during the entire study period.

Water: Animals had free access to sterile drinking water.

Cage identification: the identification labels for each cage contain the following information: number of animals, sex, strain, date received, treatment, study number, group number, and the starting date of the treatment. Animal identification: Animals were marked by ear coding.

In MDX133 Mini-PDX models, Paclitaxel treatment (T/C%=<NUM>%) could result in a decrease in tumor cell viability, Irinotecan treatment (T/C%=<NUM>%), S-<NUM> treatment (T/C%=<NUM>%), Oxaliplatin treatment (T/C%=<NUM>%) also shown a little anti-tumor activity; <NUM>-Fu treatment (T/C%=<NUM>%) shown no anti-tumor activity.

In MDX154 Mini-PDX models, all the treatment group include Gemcitabine treatment (T/C%=<NUM>%), Oxaliplatine treatment (T/C%=<NUM>%), <NUM>-Fu treatment (T/C%=<NUM>%), Nab-paclitaxel treatment (T/C%=<NUM>%), Irinotecan treatment (T/C%=<NUM>%) shown no anti-tumor activity.

In MDX164 Mini-PDX models, Temozolomide treatment (T/C%=<NUM>%) shown no anti-tumor activity.

In MDX165 Mini-PDX models, Temozolomide treatment (T/C%=<NUM>%) could result in a decrease in tumor cell viability.

In MDX168 Mini-PDX models, Nab-paclitaxel treatment (T/C%=<NUM>%), Irinotecan treatment (T/C%=<NUM>%) could result in a decrease in tumor cell viability; Gemcitabine treatment (T/C%=<NUM>%), Oxaliplatin treatment (T/C%=<NUM>%) also shown a little anti-tumor activity; <NUM>-Fu treatment (T/C%=<NUM>%) shown no anti-tumor activity.

In MDX169 Mini-PDX models, Docetaxel and Carboplatin combination treatment (T/C%=<NUM>%), Paclitaxel and Carboplatin combination treatment (T/C%=<NUM>%) could result in a decrease in tumor cell viability; Pemetrexed and Carboplatin combination treatment (T/C%=<NUM>%), Gemcitabine and Carboplatin combination treatment (T/C%=<NUM>%), Vinorelbine and Carboplatin combination treatment (T/C%=<NUM> %) also shown a little anti-tumor activity.

In MDXI <NUM> Mini-PDX models, Gemcitabine and Carboplatin combination treatment (T/C%=<NUM>%) shown a little anti-tumor activity; Pemetrexed and Carboplatin combination treatment (T/C%=<NUM>%), Paclitaxel and Carboplatin combination treatment (T/C%=<NUM>%), Docetaxel and Carboplatin combination treatment (T/C%=<NUM>%), Vinorelbine and Carboplatin combination treatment (T/C%=<NUM>%) shown no anti-tumor activity.

In MDX186 Mini-PDX models, Nab-paclitaxel treatment (T/C%=<NUM>%), Irinotecan treatment (T/C%=<NUM>%) could result in a significant decrease in tumor cell viability; Oxaliplatin treatment (T/C%=<NUM>%) shown a anti-tumor activity; Gemcitabine treatment (T/C%=<NUM>%), <NUM>-Fu treatment (T/C%=<NUM>%) also shown a little anti-tumor activity.

In MDX189 Mini-PDX models, S-<NUM> and Oxaliplatin combination treatment (T/C%=-<NUM>%), Docetaxel, Cisplatin and <NUM>-Fu combination treatment (T/C%=<NUM>% ), <NUM>-Fu and Cisplatin combination treatment (T/C%=<NUM>%) could result in a significant decrease in tumor cell viability.

In MDX203 Mini-PDX models, Oxaliplatin treatment (T/C%=<NUM>%), Irinotecan treatment (T/C%=<NUM>%), Nab-paclitaxel treatment (T/C%=<NUM>%) could result in a decrease in tumor cell viability; Gemcitabine treatment (T/C%=<NUM>%), <NUM>-Fu treatment (T/C%=<NUM>%) also showed a little anti-tumor activity.

Tumor cells numbers are counted using methods known in the art and then converted to tumor cell concentration in the suspension. M: *<NUM><NUM>/mL.

Claim 1:
A method for obtaining an animal model for screening anti-tumor drugs, comprising
(<NUM>) cultivating a primary tumor cell obtained from a tumor sample of a patient in a composition comprising:
a) <NUM>/L Hydrocortisone;
b) <NUM>/L Insulin;
c) <NUM>µg/L Cholera toxin;
d) <NUM>/L Adenine;
e) <NUM>µg/L EGF;
f) <NUM>µmol/L Y- <NUM>;
g) Pen/Strep;
h) <NUM>% FBS;
i) F12 medium
j) DMEM with high glucose
k) Non-Essential Amino Acids Solution;
l) GlutaMAX Supplement; and
m) a mouse embryonic fibroblast cell that has been treated with mitomycin C; and
(<NUM>) implanting the primary tumor cell obtained from step (<NUM>) into an animal, wherein the animal is a mouse.