CN110117527A - A kind of reinforcing discharge method of stem cell metabolic waste - Google Patents
A kind of reinforcing discharge method of stem cell metabolic waste Download PDFInfo
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- CN110117527A CN110117527A CN201910403670.0A CN201910403670A CN110117527A CN 110117527 A CN110117527 A CN 110117527A CN 201910403670 A CN201910403670 A CN 201910403670A CN 110117527 A CN110117527 A CN 110117527A
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- 210000000130 stem cell Anatomy 0.000 title claims abstract description 183
- 239000002699 waste material Substances 0.000 title claims abstract description 113
- 230000002503 metabolic effect Effects 0.000 title claims abstract description 106
- 238000000034 method Methods 0.000 title claims abstract description 33
- 230000003014 reinforcing effect Effects 0.000 title claims abstract description 27
- 239000012528 membrane Substances 0.000 claims abstract description 29
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims abstract description 29
- 235000016709 nutrition Nutrition 0.000 claims abstract description 13
- 230000035764 nutrition Effects 0.000 claims abstract description 13
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 38
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical class CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 26
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 24
- 229910021529 ammonia Inorganic materials 0.000 claims description 19
- 239000001569 carbon dioxide Substances 0.000 claims description 12
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 12
- 235000015097 nutrients Nutrition 0.000 claims description 9
- 239000004310 lactic acid Substances 0.000 claims description 7
- 235000014655 lactic acid Nutrition 0.000 claims description 7
- 239000000835 fiber Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 230000002572 peristaltic effect Effects 0.000 claims description 6
- 238000004064 recycling Methods 0.000 claims description 2
- 231100000033 toxigenic Toxicity 0.000 description 10
- 230000001551 toxigenic effect Effects 0.000 description 10
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 8
- 230000019522 cellular metabolic process Effects 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 239000004408 titanium dioxide Substances 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 239000003513 alkali Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000002253 acid Substances 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
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- 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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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Abstract
A kind of reinforcing discharge method of stem cell metabolic waste, the present invention relates to the discharge methods of stem cell metabolic waste.Solve the problem of that stem cell metabolic waste can not be discharged in time in the extensive 3D culture systems of existing stem cell.Method: one, culture solution containing stem cell metabolic waste is introduced into buffer container；Two, membrane module is selected；Three, pressurized circulation system makes the culture solution containing stem cell metabolic waste by the specific selection in membrane module through film, then the culture solution for removing stem cell metabolin is returned in buffer container, the stem cell metabolic waste isolated is transported in recyclable device by the discharge system of stem cell metabolic waste；Four, nutrition composition is added to the culture solution of removing stem cell metabolin, subsequently into stem cell incubator.The present invention is used for the reinforcing discharge method of stem cell metabolic waste.
Description
Technical field
The present invention relates to the discharge methods of stem cell metabolic waste.
Background technique
In recent years, international, domestic stem cell market development is rapid, and stem cell clinical treatment is done by public recognition and acceptance
Cell requirements are constantly promoted.Stem cell culture develops from 2D to 3D, makes great efforts the output for expanding stem cell.In the big rule of stem cell
In mould 3D culture systems, sufficiently grow in order to which stem cell is effectively ensured it is necessary that in cultivating system (stem cell incubator)
Stem cell metabolic waste must be discharged in time, otherwise just will affect stem cell growth, even cause stem cell dead.Pass through merely
Replacement culture solution is able to solve the discharge of stem cell metabolic waste, but will cause the huge waste of culture solution simultaneously.
Summary of the invention
The invention solves in the extensive 3D culture systems of existing stem cell, stem cell metabolic waste can not be discharged in time
The problem of, and a kind of reinforcing discharge method of stem cell metabolic waste is provided.
A kind of reinforcing discharge method of stem cell metabolic waste is to sequentially include the following steps:
One, the culture solution containing stem cell metabolic waste is introduced into buffer container；
In the culture solution containing stem cell metabolic waste, stem cell metabolic waste is ammonia, lactic acid class and titanium dioxide
One of carbon or in which several mixing；
When the described stem cell metabolic waste when containing ammonia, the concentration of ammonia is greater than in the culture solution containing stem cell metabolic waste
3mmol/L；
When the stem cell metabolic waste class containing lactic acid, lactic acid class in the culture solution containing stem cell metabolic waste
Concentration is greater than 24.8mmol/L；
When the stem cell metabolic waste carbonated, titanium dioxide in the culture solution containing stem cell metabolic waste
The concentration of carbon is greater than 40%；
Two, membrane module is selected；
Three, the discharge system of pressurized circulation system and stem cell metabolic waste is established, pressurized circulation system makes containing dry
The culture solution of cell metabolism waste penetrates film by the specific selection in membrane module, then will remove the training of stem cell metabolin
Nutrient solution returns in buffer container, and the stem cell metabolic waste isolated is transported to back by the discharge system of stem cell metabolic waste
In receiving apparatus；
Four, nutrition composition is added to the culture solution of removing stem cell metabolin, it is subsequently into stem cell incubator, i.e., complete
At a kind of reinforcing discharge method of stem cell metabolic waste.
The beneficial effects of the present invention are:
The culture solution containing stem cell metabolic waste is placed in a buffer container using the present invention, it will using membrane technology
Stem cell metabolic waste in culture solution selectively removes, then the nutrition composition of stem cell culture is added into culture solution, circulation
Using stem cell medium, guarantee the reinforcing discharge of stem cell metabolic waste, culture solution dosage reduces 50%~75%, effectively drops
Low stem cell toxigenic capacity 30%~75%.
The present invention is used for a kind of reinforcing discharge method of stem cell metabolic waste.
Specific embodiment
Specific embodiment 1: a kind of reinforcing discharge method of stem cell metabolic waste of present embodiment is according to the following steps
It carries out:
One, the culture solution containing stem cell metabolic waste is introduced into buffer container；
In the culture solution containing stem cell metabolic waste, stem cell metabolic waste is ammonia, lactic acid class and titanium dioxide
One of carbon or in which several mixing；
When the described stem cell metabolic waste when containing ammonia, the concentration of ammonia is greater than in the culture solution containing stem cell metabolic waste
3mmol/L；
When the stem cell metabolic waste class containing lactic acid, lactic acid class in the culture solution containing stem cell metabolic waste
Concentration is greater than 24.8mmol/L；
When the stem cell metabolic waste carbonated, titanium dioxide in the culture solution containing stem cell metabolic waste
The concentration of carbon is greater than 40%；
Two, membrane module is selected；
Three, the discharge system of pressurized circulation system and stem cell metabolic waste is established, pressurized circulation system makes containing dry
The culture solution of cell metabolism waste penetrates film by the specific selection in membrane module, then will remove the training of stem cell metabolin
Nutrient solution returns in buffer container, and the stem cell metabolic waste isolated is transported to back by the discharge system of stem cell metabolic waste
In receiving apparatus；
Four, nutrition composition is added to the culture solution of removing stem cell metabolin, it is subsequently into stem cell incubator, i.e., complete
At a kind of reinforcing discharge method of stem cell metabolic waste.
The culture solution that present embodiment removes stem cell metabolin returns in buffer container, after adding nutrition composition
Nutrition is provided into stem cell incubator for stem cell.
Present embodiment completes the discharge of specific stem cell metabolic waste using the permselective membrane of specificity.
The reinforcing discharge method of present embodiment stem cell metabolic waste is applied to stem cell and cultivates field.
Present embodiment makes the stem cell metabolic waste in stem cell medium selectively using membrane separation technique
Discharge, while retaining the nutrition composition in stem cell medium (function ingredients) recycling, the culture of stem cell is effectively reduced
Cost；Core is to select optimum seperation film for stem cell metabolin.
The beneficial effect of present embodiment is: using present embodiment by the training containing stem cell metabolic waste
Nutrient solution is placed in a buffer container, is selectively removed the stem cell metabolic waste in culture solution using membrane technology, then Xiang Pei
The nutrition composition of stem cell culture is added in nutrient solution, stem cell medium is recycled, and guarantees the reinforcing of stem cell metabolic waste
Discharge, culture solution dosage reduce 50%~75%, stem cell toxigenic capacity 30%~75% are effectively reduced.
Specific embodiment 2: the present embodiment is different from the first embodiment in that: it is special described in step 3
Property permselective membrane be hollow-fibre membrane or rolled film.It is other same as the specific embodiment one.
Specific embodiment 3: unlike one of present embodiment and specific embodiment one or two: institute in step 3
The concentration of ammonia is less than 2mmol/L in the culture solution for the removing stem cell metabolin stated；Removing stem cell generation described in step 3
The concentration for thanking to lactic acid class in the culture solution of object is less than 24.8mmol/L；The culture of stem cell metabolin is removed described in step 3
The concentration of carbon dioxide is less than 40% in liquid.It is other the same as one or two specific embodiments.
Specific embodiment 4: unlike one of present embodiment and specific embodiment one to three: institute in step 3
The pressurized circulation system stated is the pressurized circulation system that control valve and peristaltic pump cooperatively form.It is other with specific embodiment one to
Three is identical.
Specific embodiment 5: unlike one of present embodiment and specific embodiment one to four: institute in step 3
The recyclable device stated is the recyclable device equipped with recovered liquid.It is other identical as specific embodiment one to four.
Beneficial effects of the present invention are verified using following embodiment:
Embodiment one:
A kind of reinforcing discharge method of stem cell metabolic waste is to sequentially include the following steps:
One, in sterile clean culture environment, the culture solution containing stem cell metabolic waste is introduced into buffer container；
The stem cell metabolic waste is ammonia, and the concentration of ammonia is greater than in the culture solution containing stem cell metabolic waste
3mmol/L；
Two, membrane module is selected；
Three, the discharge system of pressurized circulation system and stem cell metabolic waste is established, pressurized circulation system makes containing dry
The culture solution of cell metabolism waste penetrates film by the specific selection in membrane module, then will remove the training of stem cell metabolin
Nutrient solution returns in buffer container, and the stem cell metabolic waste isolated is transported to back by the discharge system of stem cell metabolic waste
In receiving apparatus；
Four, nutrition composition is added to the culture solution of removing stem cell metabolin, it is subsequently into stem cell incubator, i.e., complete
At a kind of reinforcing discharge method of stem cell metabolic waste.
The concentration that ammonia in the culture solution of stem cell metabolin is removed described in step 3 is less than 2mmol/L；
Specific selection described in step 3 is the hollow-fibre membrane for removing ammonia through film；It is special described in step 3
Property permselective membrane purchase scientific and technological (Xiamen) Co., Ltd at the beginning of state；
Pressurized circulation system described in step 3 is the pressurized circulation system that control valve and peristaltic pump cooperatively form.
Recyclable device described in step 2 is the recyclable device equipped with acid recovery liquid.
The present embodiment method needs to keep system sterile.
The present embodiment effectively improves the service efficiency of stem cell medium, conventional stem cell medium dosage from 15 liters/
Batch is reduced to 4 liters/batch, and stem cell toxigenic capacity is effectively reduced, and the toxigenic capacity of 70,000,000 stem cells is dropped from about 80,000
Down to 40,000.
Embodiment two:
One, in sterile clean culture environment, the culture solution containing stem cell metabolic waste is introduced into buffer container；
The stem cell metabolic waste is lactic acid, and the concentration of lactic acid is greater than in the culture solution containing stem cell metabolic waste
30mmol/L；
Two, membrane module is selected；
Three, the discharge system of pressurized circulation system and stem cell metabolic waste is established, pressurized circulation system makes containing dry
The culture solution of cell metabolism waste penetrates film by the specific selection in membrane module, then will remove the training of stem cell metabolin
Nutrient solution returns in buffer container, and the stem cell metabolic waste isolated is transported to back by the discharge system of stem cell metabolic waste
In receiving apparatus；
Four, nutrition composition is added to the culture solution of removing stem cell metabolin, it is subsequently into stem cell incubator, i.e., complete
At a kind of reinforcing discharge method of stem cell metabolic waste.
The concentration that lactic acid class in the culture solution of stem cell metabolin is removed described in step 3 is less than 24.8mmol/L；
Specific selection described in step 3 is the rolled film for removing lactic acid through film；Specificity described in step 3
Permselective membrane purchase scientific and technological (Xiamen) Co., Ltd at the beginning of state；
Pressurized circulation system described in step 3 is the pressurized circulation system that control valve and peristaltic pump cooperatively form.
The present embodiment method needs to keep system sterile.
The present embodiment effectively improves the service efficiency of stem cell medium, conventional stem cell medium dosage from 14 liters/
Batch is reduced to 4 liters/batch, is effectively reduced stem cell toxigenic capacity, and the toxigenic capacities of 70,000,000 stem cells is from about 6~8
Ten thousand are reduced to 1.8~2.4 ten thousand.
Embodiment three:
A kind of reinforcing discharge method of stem cell metabolic waste is to sequentially include the following steps:
One, in sterile clean culture environment, the culture solution containing stem cell metabolic waste is introduced into buffer container；
The stem cell metabolic waste is carbon dioxide, carbon dioxide in the culture solution containing stem cell metabolic waste
Concentration is greater than 40%；
Two, membrane module is selected；
Three, the discharge system of pressurized circulation system and stem cell metabolic waste is established, pressurized circulation system makes containing dry
The culture solution of cell metabolism waste penetrates film by the specific selection in membrane module, then will remove the training of stem cell metabolin
Nutrient solution returns in buffer container, and the stem cell metabolic waste isolated is transported to back by the discharge system of stem cell metabolic waste
In receiving apparatus；
Four, nutrition composition is added to the culture solution of removing stem cell metabolin, it is subsequently into stem cell incubator, i.e., complete
At a kind of reinforcing discharge method of stem cell metabolic waste.
The concentration of carbon dioxide in the culture solution of stem cell metabolin is removed described in step 3 less than 40%；
Specific selection described in step 3 is the hollow-fibre membrane of carbon dioxide removal through film；Described in step 3
Specific selection buy scientific and technological (Xiamen) Co., Ltd at the beginning of state through film；
Pressurized circulation system described in step 2 is the pressurized circulation system that control valve and peristaltic pump cooperatively form.
Recyclable device described in step 3 is the recyclable device equipped with alkali collection liquid.
The present embodiment method needs to keep system sterile.
The present embodiment effectively improves the service efficiency of stem cell medium, conventional stem cell medium dosage from 15 liters/
Batch is reduced to 5 liters/batch, and stem cell toxigenic capacity is effectively reduced, and the toxigenic capacity of 70,000,000 stem cells is left from about 60,000
The right side is reduced to 20,000 or so.
Example IV:
A kind of reinforcing discharge method of stem cell metabolic waste is to sequentially include the following steps:
One, in sterile clean culture environment, the culture solution containing stem cell metabolic waste is introduced into buffer container；
The stem cell metabolic waste is the mixing of ammonia, lactic acid class and carbon dioxide, contains stem cell metabolic waste
The concentration of ammonia is greater than 3mmol/L in culture solution；The concentration of lactic acid class is greater than in culture solution containing stem cell metabolic waste
30mmol/L；The concentration of carbon dioxide is greater than 40% in culture solution containing stem cell metabolic waste；
Two, membrane module is selected；
Three, the discharge system of pressurized circulation system and stem cell metabolic waste is established, pressurized circulation system makes containing dry
The culture solution of cell metabolism waste penetrates film by the specific selection in membrane module, then will remove the training of stem cell metabolin
Nutrient solution returns in buffer container, and the stem cell metabolic waste isolated is transported to back by the discharge system of stem cell metabolic waste
In receiving apparatus；
Four, nutrition composition is added to the culture solution of removing stem cell metabolin, it is subsequently into stem cell incubator, i.e., complete
At a kind of reinforcing discharge method of stem cell metabolic waste.
The concentration that ammonia in the culture solution of stem cell metabolin is removed described in step 3 is less than 2mmol/L；In step 3
The concentration of lactic acid class is less than 24.8mmol/L in the culture solution of the removing stem cell metabolin；Removing described in step 3
The concentration of carbon dioxide is less than 40% in the culture solution of stem cell metabolin；
Specific selection described in step 2 through film be remove ammonia hollow-fibre membrane, removing lactic acid rolled film and
The hollow-fibre membrane of carbon dioxide removal；Specific selection described in step 2, which buys scientific and technological at the beginning of state (Xiamen) through film, to be had
Limit company；
Pressurized circulation system described in step 2 is the pressurized circulation system that control valve and peristaltic pump cooperatively form.
Recyclable device described in step 3 is the recyclable device equipped with alkali collection liquid.
The present embodiment method needs to keep system sterile.
The present embodiment effectively improves the service efficiency of stem cell medium, conventional stem cell medium dosage from 15 liters/
Batch is reduced to 4 liters/batch, and stem cell toxigenic capacity is effectively reduced, and the toxigenic capacity of 70,000,000 stem cells is left from about 60,000
The right side is reduced to 1.7 ten thousand or so.
Claims (5)
1. a kind of reinforcing discharge method of stem cell metabolic waste, it is characterised in that a kind of reinforcing discharge of stem cell metabolic waste
Method is to sequentially include the following steps:
One, the culture solution containing stem cell metabolic waste is introduced into buffer container；
In the culture solution containing stem cell metabolic waste, stem cell metabolic waste is in ammonia, lactic acid class and carbon dioxide
One kind or in which several mixing；
When the described stem cell metabolic waste when containing ammonia, the concentration of ammonia is greater than in the culture solution containing stem cell metabolic waste
3mmol/L；
When the stem cell metabolic waste class containing lactic acid, the concentration of lactic acid class in the culture solution containing stem cell metabolic waste
Greater than 30mmol/L；
When the stem cell metabolic waste carbonated, carbon dioxide in the culture solution containing stem cell metabolic waste
Concentration is greater than 40%；
Two, membrane module is selected；
Three, the discharge system of pressurized circulation system and stem cell metabolic waste is established, pressurized circulation system makes containing stem cell
The culture solution of metabolic waste penetrates film by the specific selection in membrane module, then will remove the culture solution of stem cell metabolin
Back in buffer container, the stem cell metabolic waste isolated is transported to recycling dress by the discharge system of stem cell metabolic waste
In setting；
Four, nutrition composition is added to the culture solution of removing stem cell metabolin, subsequently into stem cell incubator, that is, completes one
The reinforcing discharge method of kind stem cell metabolic waste.
2. a kind of reinforcing discharge method of stem cell metabolic waste according to claim 1, it is characterised in that in step 3
The specific selection is hollow-fibre membrane or rolled film through film.
3. a kind of reinforcing discharge method of stem cell metabolic waste according to claim 1, it is characterised in that in step 3
The concentration of ammonia is less than 2mmol/L in the culture solution of the removing stem cell metabolin；Removing stem cell described in step 3
The concentration of lactic acid class is less than 24.8mmol/L in the culture solution of metabolin；The training of stem cell metabolin is removed described in step 3
The concentration of carbon dioxide is less than 40% in nutrient solution.
4. a kind of reinforcing discharge method of stem cell metabolic waste according to claim 1, it is characterised in that in step 3
The pressurized circulation system is the pressurized circulation system that control valve and peristaltic pump cooperatively form.
5. a kind of reinforcing discharge method of stem cell metabolic waste according to claim 1, it is characterised in that in step 3
The recyclable device is the recyclable device equipped with recovered liquid.
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| WO2023012523A1 (en) * | 2019-12-02 | 2023-02-09 | Avant Meats Company Limited | System for producing cultivated meats, tissues and associated products from cells |
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