Method for dynamically and accurately detecting immunologic characteristics of primary tumor cells

A technology of primary tumor cells and detection methods, which can be used in the detection of immune characteristics of primary tumor cells and the prediction of the effect of immunotherapy in cancer patients, which can solve problems such as limited effect.

Pending Publication Date: 2021-06-18
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these measures have been found to be somewhat relevant to patient outcomes in retrospective analyses, their role remains limited in terms of predictive markers

Method used

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  • Method for dynamically and accurately detecting immunologic characteristics of primary tumor cells
  • Method for dynamically and accurately detecting immunologic characteristics of primary tumor cells
  • Method for dynamically and accurately detecting immunologic characteristics of primary tumor cells

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Experimental program
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Embodiment Construction

[0042] 1. Materials

[0043] DMEM medium: purchased from Gibco, USA

[0044] Opti-MEM medium: purchased from Gibco, USA

[0045] F12 medium: purchased from Gibco, USA

[0046] F12K medium: purchased from Gibco, USA

[0047] 10% fetal bovine serum: purchased from Gibco, USA

[0048] Streptomycin-penicillin solution: purchased from Thermo, USA

[0049] Trypsin: purchased from Gibco, USA

[0050] PBS: Phosphate buffered saline (1×), 0.0067M (PO4)

[0051] PEI: purchased from Sigma-Aldrich

[0052] EGF: purchased from Shanghai Shenggong Company

[0053] Hydrocortisone: provided by Huashan Hospital Affiliated to Fudan University

[0054] Insulin: provided by Huashan Hospital Affiliated to Fudan University

[0055] Y-27632: purchased from Selleck, USA

[0056] Collagenase: purchased from Sigma, USA

[0057] Dispase: purchased from Gibco, USA

[0058] Hyaluronidase: purchased from Sigma, USA

[0059] Protein A immunoprecipitation magnetic beads: purchased from Cell Signa...

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Abstract

The invention belongs to the technical field of medicines, and particularly relates to a method for detecting immunologic characteristics of primary tumor cells. The in-vitro detection method for the immunologic characteristics of the primary tumor cells comprises the following steps: culturing the primary tumor cells from different patient sources, establishing a bladder cancer primary tumor cell bank, detecting the expression level of PD-L1 before and after the primary cell IFN-gamma stimulation through a high-throughput flow detection platform, incubating the primary cells and the recombinant human PD-1-Fc chimeric protein, and detecting the binding capacity of the primary cells to the PD-1 protein. The immunotherapy effect of the corresponding patient is predicted in combination with the series of indexes. According to the invention, a convenient primary cell culture method and a high-throughput flow detection platform are utilized, and the individual prognosis evaluation of the in-vitro platform on cancer patients is realized by dynamically, quickly, accurately and quantitatively detecting the immune-related protein expression level and the in-vitro binding capacity of primary tumor cells.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for detecting immune characteristics of primary tumor cells, and based on this method, the effect of immunotherapy for cancer patients is predicted. Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system, and its incidence is mainly in men. Its incidence rate ranks first among male genitourinary system malignant tumors, and it has been increasing year by year in recent years. Unlike many other cancers, bladder cancer survival rates have not improved significantly over the past three decades. Despite standard treatment options, non-muscle-invasive patients have a 5-year recurrence rate as high as 60-70% and a progression rate as high as 45%. For patients with muscle-invasive disease, the response rate of clinical chemotherapy is less than 20%, and the prognosis of patients is relatively poor, with a risk of deat...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N15/14
CPCG01N15/14G01N33/56966G01N33/68
Inventor 党永军姜帅王嘉琦李增霞蒋维朱继莹
Owner FUDAN UNIV
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