Preparation method of large-scale culture dendritic cell vaccine and application thereof

A dendritic cell, large-scale culture technology, applied in the field of large-scale cultured dendritic cell vaccine preparation, can solve problems such as affecting patient treatment

Inactive Publication Date: 2013-09-18
泰州市数康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, the source of obtaining dendritic cells is mainly to isolate mononuclear cells from the peripheral blood of patients, and the collection of peripheral blood of patients is limited, generally 50 ml to 300 ml, so that the obtained dendritic

Method used

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  • Preparation method of large-scale culture dendritic cell vaccine and application thereof
  • Preparation method of large-scale culture dendritic cell vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Large scale culture of dendritic cell vaccine from mononuclear cells separated by blood cell separator

[0031] Gastric cancer patient, male, 35 years old, blood routine test result was 4.2×10 9 white blood cells per liter, 0.5L peripheral blood was collected by a blood cell separator, and the number of collected white blood cells was 2.1×10 9 (Sign an informed consent form with the patient); resuspend the collected white blood cells with 100ml 1×PBS (pH=7.4), add 25ml of 0.6% hydroxyethyl starch saline, mix well and let stand for 30 minutes, absorb as much as possible White cell layer; the white cell layer was subjected to Ficoll-Hypaque density gradient centrifugation to obtain mononuclear cells resuspended in RPMI1640 medium, and the trypan blue staining count was 1.44×10 9 .

[0032] Peripheral blood mononuclear cells were adjusted to a cell concentration of 5×10 with RPMI1640 medium 6 , added to a 2-layer cell culture device (Corning) to adhere to the wall, at 3...

Embodiment 2

[0037] Large-scale culture of dendritic cells obtained from mononuclear cells collected by apheresis

[0038] Patient 1: lung cancer patient, male, 43 years old, blood routine test result was 2.8×10 9 white blood cells per liter, using a blood cell separator to collect 1L of peripheral blood, and the number of collected white blood cells was 2.18×10 9 .

[0039] Patient 2: Breast cancer patient, female, 51 years old, blood routine test result was 3.2×10 9 white blood cells per liter, 0.75L peripheral blood was collected by a blood cell separator, and the number of collected white blood cells was 2.4×10 9 .

[0040] Patient 3: Esophageal cancer patient, male, 31 years old, blood routine test result was 7.0×10 9 white blood cells per liter, 0.5L of peripheral blood was collected using a blood cell separator, and the number of collected white blood cells was 3.5×10 9 .

[0041] The existing technology directly collects the patient's peripheral blood for the cultivation of d...

Embodiment 3

[0051] Clinical experiment of dendritic cell vaccine obtained by large-scale culture of the present invention

[0052] Clinical experiments were performed using the dendritic cell vaccine prepared in Example 2 of the present invention.

[0053] The patient's dendritic cell vaccine obtained by large-scale culture and the patient's dendritic cell vaccine obtained by direct peripheral blood mononuclear cell culture of the present invention are clinically applied to the patient (the informed consent is signed with the patient). The patient received four dendritic cell vaccine treatments in one course of treatment, once every week, and each time the dendritic cells were 5×10 6 , Intradermal injection in the abdominal groove lymph node site. Among them, dendritic cell vaccines obtained through large-scale culture can be used for multiple courses of treatment, and the next course of treatment is performed every two months, a total of 2.5 to 3 courses of treatment; while patients who...

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Abstract

The invention provides a preparation method of a large-scale culture dendritic cell vaccine. The preparation method comprises the following process steps of: performing in-vitro separation and purification on collected white blood cells in human peripheral blood by adopting a blood cell separation machine to obtain a large number of mononuclear cells, wherein the order of magnitude can be above 10<9>; using a human granulocyte-macrophage colony stimulating factor and interleukin 4 to induce the culture to obtain a large number of immature dendritic cells, wherein the order of magnitude can be above 10<8>; and loading a tumor specific antigen in the immature dendritic cells, and then obtaining the mature dendritic cell vaccine with an anti-tumor effect under the induction culture conditions of a tumor necrosis factor alpha and lipopolysaccharides. The invention further provides a multi-time and multi-treatment course clinical application of the dendritic cell vaccine obtained through the method.

Description

technical field [0001] The invention relates to a method for preparing a large-scale cultured dendritic cell vaccine. The invention relates to a large amount of mononuclear cells obtained by collecting leukocytes in human peripheral blood by adopting a blood cell separator and separating and purifying them in vitro. The obtained mononuclear cells can easily obtain a large number of dendritic cells, which highly express co-stimulatory molecules and have the function of activating the body's effector T lymphocytes, and can be used for multiple cancers including solid tumors and blood tumors course of immunotherapy. Background technique [0002] Cellular immunotherapy for cancer has been widely used clinically. It has no huge toxic and side effects caused by radiotherapy and chemotherapy, and achieves a relatively safe and dignified treatment that improves the quality of life of patients. Dendritic cells are the largest antigen-presenting cells in the body and the body's effe...

Claims

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Application Information

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IPC IPC(8): A61K39/00C12N5/078C12N5/0784A61P35/00
Inventor 宋现让王俊荣
Owner 泰州市数康生物科技有限公司
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