WO2023060614A1 - Cell culture dish for peripheral blood mononuclear cell culture - Google Patents

Cell culture dish for peripheral blood mononuclear cell culture Download PDF

Info

Publication number
WO2023060614A1
WO2023060614A1 PCT/CN2021/124265 CN2021124265W WO2023060614A1 WO 2023060614 A1 WO2023060614 A1 WO 2023060614A1 CN 2021124265 W CN2021124265 W CN 2021124265W WO 2023060614 A1 WO2023060614 A1 WO 2023060614A1
Authority
WO
WIPO (PCT)
Prior art keywords
cell culture
peripheral blood
dish
blood mononuclear
culture dish
Prior art date
Application number
PCT/CN2021/124265
Other languages
French (fr)
Chinese (zh)
Inventor
孟永辉
Original Assignee
南通市巨久新材料科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 南通市巨久新材料科技有限公司 filed Critical 南通市巨久新材料科技有限公司
Priority to PCT/CN2021/124265 priority Critical patent/WO2023060614A1/en
Priority to DE212021000177.7U priority patent/DE212021000177U1/en
Publication of WO2023060614A1 publication Critical patent/WO2023060614A1/en

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/10—Petri dish
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/20—Material Coatings

Definitions

  • the utility model relates to a cell culture dish used for culturing peripheral blood mononuclear cells.
  • Peripheral blood mononuclear cells are cells with a single nucleus in peripheral blood, including lymphocytes, monocytes, phagocytes, and dendritic cells. Peripheral blood mononuclear cells are often used in the research of autoimmune diseases, malignant tumors, organ transplantation and drug sensitivity tests, and their applications are becoming more and more mature.
  • the utility model aims to provide a cell culture dish for culturing peripheral blood mononuclear cells.
  • a cell culture dish for culturing peripheral blood mononuclear cells comprising a dish body and a dish cover adapted to the dish body; a coating layer is provided on the inner wall of the dish body, and the coating layer is made of bellflower polysaccharide material, The thickness of the coating layer is 0.5-0.7mm.
  • both the vessel body and the vessel cover are made of glass.
  • the culture dish provided by the utility model can effectively promote the expansion of peripheral blood mononuclear cells in vitro.
  • Fig. 1 is a schematic diagram of a cell culture dish for promoting the in vitro expansion of peripheral blood mononuclear cells, wherein 1 is a dish body, 2 is a dish cover, and the coating layer is not drawn.
  • Example 1 Preparation of a cell culture dish for promoting the in vitro expansion of peripheral blood mononuclear cells
  • Step S1 dissolving platycodon polysaccharide (sterilized by gamma ray cobalt 60, purchased from Sichuan Weikeqi Biotechnology Co., Ltd., number: wkq-13106) in sterile water to prepare platycodon polysaccharide with a concentration of 3 mg/mL solution, spare;
  • Step S2 taking the above-mentioned Platycodon grandiflorum polysaccharide solution to fill up the petri dish, and standing overnight in a refrigerator at 4°C;
  • Step S3 after slowly pouring the solution in the culture dish, drying at 50° C. for 1 hour, a platycodon polysaccharide coating layer can be formed on the inner wall of the culture dish, and the thickness of the coating layer is 0.5-0.7 mm.
  • the cells in the centrifuge tube are divided into four layers from top to bottom: the first layer is the plasma layer, the second layer is the ring-shaped milky white lymphocyte layer, the third layer is the transparent separation liquid layer, and the fourth layer is the red blood cell layer. Collect the second layer of cells and mix well with normal saline, then centrifuge at 400g (about 1500 rpm) for 20 minutes to collect the precipitate, and wash the precipitate twice to obtain peripheral blood mononuclear cells.
  • control group Peripheral blood mononuclear cells in good growth state were divided into control group and experimental group. Both the control group and the experimental group were cultured in GT-T551 medium containing 10% fetal bovine serum and 1% double antibody, and operated synchronously. The only difference was: The control group was cultured using a conventional glass culture dish (90 mm in diameter), and the experimental group was cultured using the cell culture dish prepared in Example 1.
  • the in vitro expansion efficiency was determined by cell counting. Before culture, the concentration of peripheral blood mononuclear cells in the control group and the experimental group were both 5 ⁇ 10 4 cells/mL; after 72 hours of culture, the concentration of peripheral blood mononuclear cells in the control group was 1.2 ⁇ 10 The concentration of blood mononuclear cells was 2.35 ⁇ 10 5 cells/mL. The control group was amplified by 2.4 times, and the experimental group was amplified by 4.7 times.
  • the culture dish provided by the utility model can effectively promote the expansion of peripheral blood mononuclear cells in vitro.

Landscapes

  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Clinical Laboratory Science (AREA)
  • Immunology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Provided in the present invention is a cell culture dish for peripheral blood mononuclear cell culture, comprising a dish body and a dish cover fit to the dish body; an inner wall of the dish body is provided with a coating layer, the coating layer being made of Platycodon polysaccharide, and the thickness of the coating layer being 0.5-0.7 mm. The described culture dish can effectively promote the expansion of peripheral blood mononuclear cells in vitro.

Description

一种用于外周血单个核细胞培养的细胞培养皿A cell culture dish for culturing peripheral blood mononuclear cells 技术领域technical field
本实用新型涉及一种用于外周血单个核细胞培养的细胞培养皿。The utility model relates to a cell culture dish used for culturing peripheral blood mononuclear cells.
背景技术Background technique
外周血单个核细胞是外周血中具有单个核的细胞，包括淋巴细胞、单核细胞、吞噬细胞和树突状细胞等。外周血单个核细胞常用于自身免疫疾病、恶性肿瘤、器官移植和药物敏感试验等领域的研究，应用越来越成熟。Peripheral blood mononuclear cells are cells with a single nucleus in peripheral blood, including lymphocytes, monocytes, phagocytes, and dendritic cells. Peripheral blood mononuclear cells are often used in the research of autoimmune diseases, malignant tumors, organ transplantation and drug sensitivity tests, and their applications are becoming more and more mature.
发明内容Contents of the invention
本实用新型旨在提供一种用于外周血单个核细胞培养的细胞培养皿。The utility model aims to provide a cell culture dish for culturing peripheral blood mononuclear cells.
本实用新型通过如下技术方案得以实现：The utility model is realized through the following technical solutions:
一种用于外周血单个核细胞培养的细胞培养皿，包括皿体和与所述皿体适配的皿盖；所述皿体的内壁设有包被层，包被层为桔梗多糖材质，包被层厚度为0.5-0.7mm。A cell culture dish for culturing peripheral blood mononuclear cells, comprising a dish body and a dish cover adapted to the dish body; a coating layer is provided on the inner wall of the dish body, and the coating layer is made of bellflower polysaccharide material, The thickness of the coating layer is 0.5-0.7mm.
优选地，所述皿体和皿盖均为玻璃材质。Preferably, both the vessel body and the vessel cover are made of glass.
本实用新型的优点：Advantage of the utility model:
本实用新型提供的培养皿可以有效促进外周血单个核细胞体外扩增。The culture dish provided by the utility model can effectively promote the expansion of peripheral blood mononuclear cells in vitro.
附图说明Description of drawings
图1为促进外周血单个核细胞体外扩增的细胞培养皿的示意图，其中1为皿体，2为皿盖，包被层未画出。Fig. 1 is a schematic diagram of a cell culture dish for promoting the in vitro expansion of peripheral blood mononuclear cells, wherein 1 is a dish body, 2 is a dish cover, and the coating layer is not drawn.
具体实施方式Detailed ways
下面结合实施例具体介绍本实用新型实质性内容。Introduce the substantive content of the utility model in detail below in conjunction with embodiment.
实施例1：促进外周血单个核细胞体外扩增的细胞培养皿的制备Example 1: Preparation of a cell culture dish for promoting the in vitro expansion of peripheral blood mononuclear cells
取常规的玻璃培养皿(直径90mm)制备本实用新型培养皿，具体步骤如下：Get conventional glass culture dish (diameter 90mm) and prepare the utility model culture dish, concrete steps are as follows:
步骤S1，将桔梗多糖(采用γ射线钴60照射灭菌，购自四川省维克奇生物科技有限公司，编号：wkq-13106)溶于无菌水中，制成浓度为3mg/mL的桔梗多糖溶液，备用；Step S1, dissolving platycodon polysaccharide (sterilized by gamma ray cobalt 60, purchased from Sichuan Weikeqi Biotechnology Co., Ltd., number: wkq-13106) in sterile water to prepare platycodon polysaccharide with a concentration of 3 mg/mL solution, spare;
步骤S2，取上述桔梗多糖溶液加满培养皿，4℃冰箱中静置过夜；Step S2, taking the above-mentioned Platycodon grandiflorum polysaccharide solution to fill up the petri dish, and standing overnight in a refrigerator at 4°C;
步骤S3，慢慢倾倒培养皿中溶液后，50℃干燥1h，即可在培养皿内壁形成桔梗多糖包被层，包被层厚度0.5-0.7mm。Step S3, after slowly pouring the solution in the culture dish, drying at 50° C. for 1 hour, a platycodon polysaccharide coating layer can be formed on the inner wall of the culture dish, and the thickness of the coating layer is 0.5-0.7 mm.
实施例2：外周血单个核细胞体外扩增Example 2: In vitro expansion of peripheral blood mononuclear cells
1、外周血单个核细胞的提取1. Extraction of peripheral blood mononuclear cells
取健康志愿者外周抗凝血，与生理盐水等体积混匀后，小心加于人外周血淋巴细胞分离 液的液面上，以400g(约1500转/分，半径15cm水平转子)离心20分钟，此时离心管中由上至下细胞分四层：第一层为血浆层，第二层为环状乳白色淋巴细胞层，第三层为透明分离液层，第四层为红细胞层。收集第二层细胞用生理盐水充分混匀后，以400g(约1500转/分)离心20分钟，收集沉淀，沉淀重复洗涤2次后即得外周血单个核细胞。Take peripheral anticoagulant blood from healthy volunteers, mix it with normal saline, carefully add it to the liquid surface of human peripheral blood lymphocyte separation medium, and centrifuge at 400g (about 1500 rpm, horizontal rotor with a radius of 15cm) for 20 minutes At this time, the cells in the centrifuge tube are divided into four layers from top to bottom: the first layer is the plasma layer, the second layer is the ring-shaped milky white lymphocyte layer, the third layer is the transparent separation liquid layer, and the fourth layer is the red blood cell layer. Collect the second layer of cells and mix well with normal saline, then centrifuge at 400g (about 1500 rpm) for 20 minutes to collect the precipitate, and wash the precipitate twice to obtain peripheral blood mononuclear cells.
2、外周血单个核细胞分组和培养2. Grouping and culture of peripheral blood mononuclear cells
将生长状态良好的外周血单个核细胞分成对照组和实验组，对照组和实验组均使用含10％胎牛血清和1％双抗的GT-T551培养基培养，同步操作，区别仅在于：对照组使用常规的玻璃培养皿(直径90mm)培养，实验组使用实施例1制备的细胞培养皿培养。Peripheral blood mononuclear cells in good growth state were divided into control group and experimental group. Both the control group and the experimental group were cultured in GT-T551 medium containing 10% fetal bovine serum and 1% double antibody, and operated synchronously. The only difference was: The control group was cultured using a conventional glass culture dish (90 mm in diameter), and the experimental group was cultured using the cell culture dish prepared in Example 1.
3、体外扩增效率测定3. Determination of in vitro amplification efficiency
用细胞计数法测定体外扩增效率。培养前，对照组和实验组外周血单个核细胞的浓度均为5×10 4个/mL；培养72h后，对照组外周血单个核细胞的浓度为1.2×10 5个/mL，实验组外周血单个核细胞的浓度为2.35×10 5个/mL。对照组扩增了2.4倍，实验组扩增了4.7倍。 The in vitro expansion efficiency was determined by cell counting. Before culture, the concentration of peripheral blood mononuclear cells in the control group and the experimental group were both 5×10 4 cells/mL; after 72 hours of culture, the concentration of peripheral blood mononuclear cells in the control group was 1.2 ×10 The concentration of blood mononuclear cells was 2.35×10 5 cells/mL. The control group was amplified by 2.4 times, and the experimental group was amplified by 4.7 times.
由此可见，本实用新型提供的培养皿可以有效促进外周血单个核细胞体外扩增。It can be seen that the culture dish provided by the utility model can effectively promote the expansion of peripheral blood mononuclear cells in vitro.
上述实施例的作用在于具体介绍本实用新型的实质性内容，但本领域技术人员应当知道，不应将本实用新型的保护范围局限于该具体实施例。The purpose of the above embodiments is to specifically introduce the substantive content of the utility model, but those skilled in the art should know that the protection scope of the utility model should not be limited to the specific examples.

Claims (2)

  1. 一种用于外周血单个核细胞培养的细胞培养皿，其特征在于：包括皿体和与所述皿体适配的皿盖；所述皿体的内壁设有包被层，包被层为桔梗多糖材质，包被层厚度为0.5-0.7mm。A cell culture dish for culturing peripheral blood mononuclear cells is characterized in that: it includes a dish body and a dish cover adapted to the dish body; the inner wall of the dish body is provided with a coating layer, and the coating layer is Platycodon grandiflorum is made of polysaccharide, and the thickness of the coating layer is 0.5-0.7mm.
  2. 根据权利要求1所述的细胞培养皿，其特征在于：所述皿体和皿盖均为玻璃材质。The cell culture dish according to claim 1, wherein both the dish body and the dish cover are made of glass.
PCT/CN2021/124265 2021-10-16 2021-10-16 Cell culture dish for peripheral blood mononuclear cell culture WO2023060614A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/CN2021/124265 WO2023060614A1 (en) 2021-10-16 2021-10-16 Cell culture dish for peripheral blood mononuclear cell culture
DE212021000177.7U DE212021000177U1 (en) 2021-10-16 2021-10-16 A cell culture dish for culturing peripheral blood mononuclear cells

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2021/124265 WO2023060614A1 (en) 2021-10-16 2021-10-16 Cell culture dish for peripheral blood mononuclear cell culture

Publications (1)

Publication Number Publication Date
WO2023060614A1 true WO2023060614A1 (en) 2023-04-20

Family

ID=79689605

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/124265 WO2023060614A1 (en) 2021-10-16 2021-10-16 Cell culture dish for peripheral blood mononuclear cell culture

Country Status (2)

Country Link
DE (1) DE212021000177U1 (en)
WO (1) WO2023060614A1 (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1709915A (en) * 2005-06-23 2005-12-21 南京大学医学院附属鼓楼医院 Polysaccharide material for promoting cell tactophily and preparation thereof
CN201695032U (en) * 2010-07-14 2011-01-05 刘军发 Microorganism culture dish
CN103320322A (en) * 2013-06-06 2013-09-25 浙江硕华医用塑料有限公司 Cell culture dish
CN106010947A (en) * 2016-08-09 2016-10-12 贵州大学 Glass culture dish for microbiological lab

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1709915A (en) * 2005-06-23 2005-12-21 南京大学医学院附属鼓楼医院 Polysaccharide material for promoting cell tactophily and preparation thereof
CN201695032U (en) * 2010-07-14 2011-01-05 刘军发 Microorganism culture dish
CN103320322A (en) * 2013-06-06 2013-09-25 浙江硕华医用塑料有限公司 Cell culture dish
CN106010947A (en) * 2016-08-09 2016-10-12 贵州大学 Glass culture dish for microbiological lab

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
LI FENGYUAN: "Research Progress on Effective Components and Biological Functions of Platycodon Platycodon", HUBEI JOURNAL OF ANIMAL AND VETERINARY SCIENCES, vol. 38, no. 4, 5 April 2017 (2017-04-05), pages 7 - 9, XP093058155, ISSN: 1007-273X, DOI: 10.16733/j.cnki.issn1007-273x.2017.04.002 *

Also Published As

Publication number Publication date
DE212021000177U1 (en) 2022-01-04

Similar Documents

Publication Publication Date Title
CN101942413B (en) Birth defect cell bank and construction method thereof
WO2019210833A1 (en) Exosome active preparation promoting endothelial cell angiogenesis and preparation method and use thereof
CN114591905B (en) Method for preparing apoptotic vesicles from human erythrocytes and application of apoptotic vesicles
CN106539821A (en) A kind of stem cell excretion body preparation and its preparation method and application
CN106906271A (en) Application of bone marrow mesenchymal stem cell-derived exocrine body and test method thereof
CN106834290A (en) A kind of circular rna and application thereof
CN107663539A (en) Circular rna circ PTGR1 purposes
CN108103019A (en) A kind of preparation method of tumour-specific gamma delta T cells
WO2023060614A1 (en) Cell culture dish for peripheral blood mononuclear cell culture
JPS63263081A (en) Separation of malignant cell from clinical substance to be examined
WO2022042739A1 (en) Method for promoting endothelial cells to produce exosomes, and exosome preparation and application thereof
WO2023060613A1 (en) Peripheral blood mononuclear cell culture dish containing coriolus versicolor mushroom polysaccharide coating layer
CN111548994B (en) Cell culture medium and method for culturing NK cells by using same
CN206744390U (en) A kind of umbilical hemopoietic stem cell cryopreservation tube
CN113082040A (en) Prostate cancer treatment product
Sung et al. Enrichment of cancer-initiating cells from colon cancer cells through porous polymeric membranes by a membrane filtration method
CN112675203A (en) Application of cell-derived exosome in preparation of biological preparation for treating asthma and/or pulmonary fibrosis
CN115678841A (en) Composition for enhancing immune suppression function of human umbilical cord mesenchymal stem cells and application
CN113073077B (en) Method for culturing clinical-grade umbilical cord blood mesenchymal stem cells by using closed system
CN112300992B (en) NK cell culture solution and multistage activated NK cell culture method
WO2023065263A1 (en) Cell culture dish for promoting in vitro amplification of adipose mesenchymal stem cells
CN111394301B (en) Application of piceatannol in increasing number of secreted exosomes of pluripotent stem cells and bioactivity
WO2023065262A1 (en) Culture dish specially used for adipose-derived mesenchymal stem cells
WO2020258637A1 (en) Kidney transplant donor-specific urine-derived cell and its dna preparation method and application thereof
CN112538459A (en) Method for separating exosome in liver cancer tissue

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21960343

Country of ref document: EP

Kind code of ref document: A1