CN105994253A - Frozen stock solution of immune cells - Google Patents
Frozen stock solution of immune cells Download PDFInfo
- Publication number
- CN105994253A CN105994253A CN201610553886.1A CN201610553886A CN105994253A CN 105994253 A CN105994253 A CN 105994253A CN 201610553886 A CN201610553886 A CN 201610553886A CN 105994253 A CN105994253 A CN 105994253A
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- CN
- China
- Prior art keywords
- stock solution
- frozen stock
- cell
- immune cells
- trehalose
- Prior art date
- Legal status (The legal status 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 status listed.)
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Biophysics (AREA)
- Physiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a frozen stock solution of immune cells. The frozen stock solution of the immune cells comprises the following components: 25 to 100 U/mL of recombinant human interleukin-2, 0.5 to 2.5 mg/mL of alpha-1,4-glucan, 0.05 to 0.2 mg/mL of trehalose, 0.1 to 0.5 mL/mL of polyethylene glycol, 0.05 to 0.15 mL/mL of propylene glycol and 0.7 to 0.9 mL/mL of a basic medium. The frozen stock solution of the immune cells disclosed by the invention has the beneficial effects of being high in safety, small in damage to the cells and capable of guaranteeing normal biological functions after the cells are recovered.
Description
Technical field
The invention belongs to technical field of cell culture, particularly relate to a kind of immunocyte frozen stock solution.
Background technology
Immune cell therapy is the novel method for the treatment of that a kind of autoimmune is anticancer, and it is to use biotechnology and biological system
The agent immunocyte to gathering from the patient carries out the defeated method returned in patient body after In vitro culture and amplification, to excite,
Enhancing body not holding any government official post's immunologic function, thus reach to treat the purpose of tumor.Cell cryopreservation is the main method of long-term cell preservation
One of, but cell cryopreservation process can change the thermodynamics of cell, chemically and physically environment, can cause cell a certain degree of
Biological injury.In order to be down to minimum by the damage of cell in cell cryopreservation, resuscitation process, not only need to optimize chemistry and temperature
Operating process, and need to add one or more freezing protective agent before frozen, remove it the most again.
The most the most frequently used freezing protective agent is dimethyl sulfoxide (DMSO), and this molecular weight of material is little, and dissolubility is big, easily
Penetration cell, can make freeze point depression, reduces intracellular and forms the chance of ice crystal, thus reduces the ice crystal damage to cell.Due to height
Concentration DMSO to cell by toxicity, it is necessary to add other liquid component, such as serum, cell culture medium, to reduce DMSO's
Concentration, reduces the injury to cell.The hyclone of the DMSO and 90% of the many employings of existing cells frozen storing liquid 10%, but due to DMSO
Content is the highest, has suitable toxicity, unfavorable to patient body, and containing substantial amounts of foreign protein in hyclone, both
There is the danger of infectious disease, be easy to again cause allergic reaction or immunologic rejection.And the cell survival that this kind of frozen stock solution is frozen
Rate and cytoactive are relatively low.
Summary of the invention
The purpose of the present invention is that and overcomes above-mentioned deficiency, it is provided that a kind of immunocyte frozen stock solution.
For reaching above-mentioned purpose, the present invention implements according to techniques below scheme:
A kind of immunocyte frozen stock solution, including following components: RhIL-2, α-Isosorbide-5-Nitrae-glucosan, trehalose, poly-second two
Alcohol, propylene glycol and basal medium.
Further, including following components: RhIL-2 25-100 U/mL, α-Isosorbide-5-Nitrae-glucosan 0.5-2.5
Mg/mL, trehalose 0.05-0.2 mg/mL, Polyethylene Glycol 0.1-0.5 mL/mL, propylene glycol 0.05-0.15 mL/mL and basis
Culture medium 0.7-0.9 mL/mL.
Further, including following components: RhIL-2 50 U/mL, α-Isosorbide-5-Nitrae-glucosan 1.5 mg/mL, sea
Algae sugar 0.1 mg/mL, Polyethylene Glycol 0.25 mL/mL, propylene glycol 0.05 mL/mL and basal medium 0.85 mL/mL.
Further, described basal medium is RPMI-1640 or GT-T551 or GT-T561 culture medium.
Compared with prior art, the invention have the benefit that
1. the present invention can substitute dimethyl sulfoxide by adding appropriate trehalose, Polyethylene Glycol and propylene glycol, it is ensured that cell
When close to freezing point, the moisture of intracellular will not crystallize, and can be effectively protected cell, and effectively reduce cytotoxicity；
2. the present invention adds α-Isosorbide-5-Nitrae-glucosan and can effectively reduce the damage of cell in frozen, recovery, and ensure that multiple
The vigor of cell proliferation after Soviet Union；
3. the present invention adds RhIL-2 and can effectively maintain the activity of cell, it is ensured that normal biological after cell recovery
Learn function.
The immunocyte frozen stock solution safety of the present invention is high, and the damage to cell is little, it is possible to normal after ensureing cell recovery
Biological function.
Detailed description of the invention
Below in conjunction with specific embodiment, the invention will be further described, the illustrative examples invented at this and explanation
It is used for explaining the present invention, but not as a limitation of the invention.
Embodiment 1
A kind of immunocyte frozen stock solution, including following components: RhIL-2 50 U/mL, α-Isosorbide-5-Nitrae-glucosan 1.5
Mg/mL, trehalose 0.1 mg/mL, Polyethylene Glycol 0.25 mL/mL, propylene glycol 0.05 mL/mL and RPMI-1640 culture medium
0.85 mL/mL。
Embodiment 2
A kind of immunocyte frozen stock solution, including following components: RhIL-2 25 U/mL, α-Isosorbide-5-Nitrae-glucosan 1.0
Mg/mL, trehalose 0.05 mg/mL, Polyethylene Glycol 0.3 mL/mL, propylene glycol 0.1 mL/mL and GT-T551 culture medium 0.9
mL/mL。
Embodiment 3
A kind of immunocyte frozen stock solution, including following components: RhIL-2 100 U/mL, α-Isosorbide-5-Nitrae-glucosan 2.0
Mg/mL, trehalose 0.15 mg/mL, Polyethylene Glycol 0.25 mL/mL, propylene glycol 0.15 mL/mL and GT-T561 culture medium 0.9
mL/mL。
Technical scheme is not limited to the restriction of above-mentioned specific embodiment, every does according to technical scheme
The technology deformation gone out, within each falling within protection scope of the present invention.
Claims (4)
1. an immunocyte frozen stock solution, it is characterised in that include following components: RhIL-2, α-Isosorbide-5-Nitrae-glucosan,
Trehalose, Polyethylene Glycol, propylene glycol and basal medium.
A kind of immunocyte frozen stock solution the most according to claim 1, it is characterised in that include following components: recombined human is white
Interleukin-2 25-100 U/mL, alpha-1,4-dextran 0.5-2.5 mg/mL, trehalose 0.05-0.2 mg/mL, Polyethylene Glycol
0.1-0.5 mL/mL, propylene glycol 0.05-0.15 mL/mL and basal medium 0.7-0.9 mL/mL.
A kind of immunocyte frozen stock solution the most according to claim 1, it is characterised in that include following components: recombined human is white
Interleukin-2 50 U/mL, alpha-1,4-dextran 1.5 mg/mL, trehalose 0.1 mg/mL, Polyethylene Glycol 0.25 mL/mL, the third two
Alcohol 0.05 mL/mL and basal medium 0.85 mL/mL.
A kind of immunocyte frozen stock solution the most according to claim 1, it is characterised in that described basal medium is RPMI-
1640 or GT-T551 or GT-T561 culture medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610553886.1A CN105994253A (en) | 2016-07-15 | 2016-07-15 | Frozen stock solution of immune cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610553886.1A CN105994253A (en) | 2016-07-15 | 2016-07-15 | Frozen stock solution of immune cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105994253A true CN105994253A (en) | 2016-10-12 |
Family
ID=57118759
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610553886.1A Pending CN105994253A (en) | 2016-07-15 | 2016-07-15 | Frozen stock solution of immune cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105994253A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107012119A (en) * | 2017-05-31 | 2017-08-04 | 东莞市保莱生物科技有限公司 | A kind of method that immunocyte is extracted from marrow |
| CN107047536A (en) * | 2016-11-22 | 2017-08-18 | 浙江三誉生物科技有限公司 | A kind of cell-preservation liquid and its application |
| CN109744226A (en) * | 2018-12-30 | 2019-05-14 | 深圳光彩生命工程技术有限公司 | A kind of immunocyte frozen stock solution |
| CN112806354A (en) * | 2021-01-18 | 2021-05-18 | 圣至同合(北京)生物科技有限公司 | Immune cell cryopreservation liquid as well as preparation method and application thereof |
| CN114615886A (en) * | 2019-08-29 | 2022-06-10 | 得克萨斯大学体系董事会 | Cell cryopreservation culture medium |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001036589A2 (en) * | 1999-11-17 | 2001-05-25 | University Of Rochester | Human ex vivo immune system |
| CN105324480A (en) * | 2013-06-28 | 2016-02-10 | 株式会社大塚制药工场 | Trehalose and dextran-containing solution for transplanting mammalian cells |
| CN105462920A (en) * | 2015-12-31 | 2016-04-06 | 北京弘润天源生物技术股份有限公司 | Method for extracting immune cells from peripheral blood |
| CN105638642A (en) * | 2016-01-27 | 2016-06-08 | 上海润泉生物技术有限公司 | Immune cell cryopreservation solution and application thereof |
-
2016
- 2016-07-15 CN CN201610553886.1A patent/CN105994253A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001036589A2 (en) * | 1999-11-17 | 2001-05-25 | University Of Rochester | Human ex vivo immune system |
| CN105324480A (en) * | 2013-06-28 | 2016-02-10 | 株式会社大塚制药工场 | Trehalose and dextran-containing solution for transplanting mammalian cells |
| CN105462920A (en) * | 2015-12-31 | 2016-04-06 | 北京弘润天源生物技术股份有限公司 | Method for extracting immune cells from peripheral blood |
| CN105638642A (en) * | 2016-01-27 | 2016-06-08 | 上海润泉生物技术有限公司 | Immune cell cryopreservation solution and application thereof |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107047536A (en) * | 2016-11-22 | 2017-08-18 | 浙江三誉生物科技有限公司 | A kind of cell-preservation liquid and its application |
| CN107012119A (en) * | 2017-05-31 | 2017-08-04 | 东莞市保莱生物科技有限公司 | A kind of method that immunocyte is extracted from marrow |
| CN109744226A (en) * | 2018-12-30 | 2019-05-14 | 深圳光彩生命工程技术有限公司 | A kind of immunocyte frozen stock solution |
| CN114615886A (en) * | 2019-08-29 | 2022-06-10 | 得克萨斯大学体系董事会 | Cell cryopreservation culture medium |
| CN112806354A (en) * | 2021-01-18 | 2021-05-18 | 圣至同合(北京)生物科技有限公司 | Immune cell cryopreservation liquid as well as preparation method and application thereof |
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Application publication date: 20161012 |