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Human-derived chordoma bone in-situ PDX model construction method and application thereof

A construction method and chordoma technology, applied in the field of human chordoma bone orthotopic PDX model construction

Active Publication Date: 2021-09-07
SHANGHAI FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the deficiencies of the prior art, the present invention creatively proposes a method for constructing a human chordoma bone in situ PDX model for the first time, and there is no information about the method for constructing a human chordoma bone in situ PDX model of the present invention and its application. See the report

Method used

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  • Human-derived chordoma bone in-situ PDX model construction method and application thereof
  • Human-derived chordoma bone in-situ PDX model construction method and application thereof
  • Human-derived chordoma bone in-situ PDX model construction method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The construction of embodiment 1 chordoma model

[0022] 1 method

[0023] (1) After surgically removing the chordoma completely, the chordoma tissue was quickly transferred to the experimenter for processing. Under sterile conditions, place the tumor tissue in normal saline, remove blood clots, necrotic tissue, fat and connective tissue on the surface of the tumor tissue, and take typical fresh tumor tissue ( figure 1 ), use tissue scissors and tweezers alternately, and store the tumor tissue in three parts;

[0024] (2) The first tumor tissue was quickly placed in a cryopreservation tube filled with RNAlater protection solution (Invitrogen, USA) under aseptic conditions, and quickly transferred to liquid nitrogen for preservation for transcriptome sequencing;

[0025] (3) The second tumor tissue was stored in formalin, embedded in paraffin, made into a wax block and sectioned, stained with hematoxylin and eosin, observed under a microscope, and circled the typical v...

Embodiment 2

[0033] The detection of embodiment 2 chordoma model

[0034] 1 method

[0035] The model constructed in Example 1 was taken and sacrificed, and the mouse tumor tissue was taken for histopathological examination and evaluation.

[0036] 2 results

[0037] The results show that chordoma tumors can be transplanted into the tibial medullary cavity of immunodeficient mice, and the tumor retains the original tissue structure characteristics. By detecting chordoma tumor markers, it can be proved that the constructed model has a high degree of simulation, and the success rate of modeling is over 70%. , indicating that the method is feasible.

Embodiment 3

[0038] In vivo passage of embodiment 3 chordoma model

[0039] 1 method

[0040] According to the model obtained by the method described in Example 1, when the tumor volume reaches 0.5~1×10 3 mm 3 chordoma tissues in the tibia were harvested using fine ophthalmic shears, the tumors were passaged in vivo, and the validity and feasibility of the model were evaluated by histopathological examination.

[0041] 2 results

[0042] The tumor markers of passaged tumors (Brachyury, Aggrecan, Collagen type II alpha 1chain) showed that after passage in immunodeficient mice, the tumor retained the original tissue structure characteristics, the results are shown in Figure 5 , Image 6 . Chordoma can grow in the tibial medullary cavity of immunodeficient mice, which shows that transplantation by this method is feasible.

[0043] Experimental results show that at least 5 passages can be performed.

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Abstract

The invention relates to the technical field of medical engineering, and particularly relates to a human-derived chordoma bone in-situ PDX model construction method and application thereof. The human-derived chordoma bone in-situ PDX model construction method comprises the steps that (1), chordoma patient tissue is selected; (2), the chordoma tissue is divided into three parts, one part is used for transcriptome sequencing, the other part is used for making a tissue chip, and the last part is implanted into the tibia medullary cavity of an immunodeficiency mouse; (3), the growth condition of the tibia tumor of the mouse is observed; (4), passage is conducted; and (5), the effectiveness and feasibility of the method are judged, and the method is used for detecting curative effect prediction and curative effect monitoring of potential new drugs. The method has the advantages that a model obtained through a chordoma construction method in the prior art cannot well simulate the disease characteristics and cannot well conduct drug screening to obtain effective drugs, the defects are overcome, and the model obtained through the construction method has the advantages of being high in simulation degree and good in feasibility.

Description

technical field [0001] The invention relates to the technical field of medical engineering, in particular to a method for constructing a human chordoma bone in situ PDX model and its application. Background technique [0002] Chordoma is a relatively rare malignant tumor of bone and nervous system. Although its growth is relatively slow, it is highly locally invasive and often causes localized pain in the affected area. Because chordoma is often adjacent to important nerve tissues such as the brainstem and spinal cord, its growth often causes compression of nerve tissues, leading to complications such as limb sensorimotor disorders, paralysis, etc., seriously affecting the quality of life of patients and even threatening their lives. In terms of treatment, chordoma is not sensitive to radiotherapy, chemotherapy and other treatment methods, and surgical resection is the main treatment method. However, due to the particularity and complexity of the anatomical structure of th...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01K67/027
CPCC12N5/0693A01K67/0271C12N2509/00A01K2227/105A01K2267/0331
Inventor 孟通尹华斌宋滇文金佳丽高健萱李勇爱
Owner SHANGHAI FIRST PEOPLES HOSPITAL
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