Construction method and application of PDX model based on osteogenic niche microenvironment modification

A construction method and microenvironment technology, applied in the field of PDX model construction, can solve the problems of low tumor formation rate, poor stability and passage, and promotion limitations, and achieve high transplantation success rate and the effect of retaining the biological characteristics of primary cells

Active Publication Date: 2021-09-24
中国人民解放军陆军特色医学中心
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Problems solved by technology

However, due to the ethical factors involved in the acquisition of fetal cartilage, the promotion of this method is also subject to certain restrictions.
Existing leukemia PDX model grafts are cell suspensions, which lack the stromal components of solid tumor grafts or the support of "hematopoietic niches" in bone marrow tissue, resulting in low tumor formation rate and poor stability and passage. Even with some improved PDX models, Its application is also limited because of its high cost or ethical factors

Method used

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  • Construction method and application of PDX model based on osteogenic niche microenvironment modification
  • Construction method and application of PDX model based on osteogenic niche microenvironment modification
  • Construction method and application of PDX model based on osteogenic niche microenvironment modification

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Experimental program
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Effect test

Embodiment 1

[0024] Preparation of bio-derived bone

[0025] Process the cancellous bone at the medullary end of the long bone of the porcine into a bone block of 1 cm×0.8 cm×0.8 cm, put the processed bone block into 30% hydrogen peroxide, soak at 38°C for 72 hours, and change the solution every 24 hours. Then soak in double-distilled water for 30 minutes, soak in ethanol for 24 hours, soak in acetone for 24 hours, soak in double-distilled water for 30 minutes, and finally dry at room temperature for 8 hours, vacuum freeze-dry, ethylene oxide gas fumigation and sterilization, and pack them in sterile plastic bags Sealed and preserved, the shape of the bio-derived niche is as follows figure 1 Shown in A and B.

Embodiment 2

[0027] Method for constructing PDX model of microenvironment modification of "osteogenesis niche"

[0028] (1) Isolation and culture of human umbilical cord mesenchymal cells (MSCs)

[0029] Take a sterile healthy fetal umbilical cord of about 4-5 cm, wash it fully with PBS, separate and remove the cord adventitia and vascular tissue, cut the obtained umbilical cord tissue into small tissue pieces, transfer to 0.1% collagenase IV, and keep at 37°C Digest, pass through a 100-mesh sieve, collect the filtrate containing cells, centrifuge at 1200r / min for 10min at room temperature, discard the supernatant, keep the precipitate; wash twice with PBS, wash with 1×10 6 / mL cell density inoculated in DMEM / F12 medium containing 10% FBS, 37°C, 5% CO 2 cultured in a saturated humidity environment. Change the cell solution after 6-7 days, discard the non-adherent cells, and change the solution every 3-4 days thereafter. After observing that the cells reached 80% confluence, they were di...

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Abstract

The invention belongs to the technical field, and particularly relates to a construction method and application of a PDX model based on osteogenesis niche microenvironment modification. The construction method comprises the following steps: placing human umbilical cord mesenchymal cells on a biological derived bone, inducing and differentiating the human umbilical cord mesenchymal cells into osteoblasts to obtain the osteoblast-loaded biological derived bone, planting the osteoblast-loaded biological derived bone under the skin of a mouse, and inoculating the planting part of the biological derived bone under the skin of the mouse with primary leukemia cells or tumor cells. A novel microenvironment modified PDX model is constructed, and a disease research model which is high in transplantation success rate, stable in passage and capable of effectively retaining the biological characteristics of primary cells is provided for research of leukemia or tumor cell drug resistance formation mechanism and drug screening.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to a construction method and application of a PDX model based on the microenvironment modification of an "osseogenic niche". Background technique [0002] Patient-Derived tumor xenograft (PDX, Patient-Derived tumor Xenograft) model is a model that can effectively study the biological characteristics of malignant tumors, drug resistance and potential sensitive drug screening. The PDX model refers to the patient's fresh tumor After the tissue is processed, it is transplanted into immunodeficient mice and grows in the microenvironment provided by the mice. This model can retain the basic characteristics of the primary tumor cells and provides a good in vivo model for studying tumors. Compared with the traditional tumor cell line transplantation model, the PDX tumor model can be closer to the cytogenetics and molecular biology characteristics of the human body itself, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027A61K49/00
CPCA01K67/0271A61K49/0008A01K2207/12A01K2217/03A01K2227/105A01K2267/0331
Inventor 曾东风贾乙付文莹
Owner 中国人民解放军陆军特色医学中心
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