CN113308433A - Method for treating hybrid cells on surface of islet in-vitro culture - Google Patents
Method for treating hybrid cells on surface of islet in-vitro culture Download PDFInfo
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- CN113308433A CN113308433A CN202110460519.8A CN202110460519A CN113308433A CN 113308433 A CN113308433 A CN 113308433A CN 202110460519 A CN202110460519 A CN 202110460519A CN 113308433 A CN113308433 A CN 113308433A
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- 210000004754 hybrid cell Anatomy 0.000 title claims abstract description 41
- 238000000338 in vitro Methods 0.000 title claims abstract description 24
- 238000000034 method Methods 0.000 title claims abstract description 24
- 210000004153 islets of langerhan Anatomy 0.000 claims abstract description 42
- JDNTWHVOXJZDSN-UHFFFAOYSA-N iodoacetic acid Chemical compound OC(=O)CI JDNTWHVOXJZDSN-UHFFFAOYSA-N 0.000 claims abstract description 36
- 239000001963 growth medium Substances 0.000 claims abstract description 32
- 210000004027 cell Anatomy 0.000 claims abstract description 21
- 238000012258 culturing Methods 0.000 claims abstract description 10
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 claims description 20
- 238000000746 purification Methods 0.000 claims description 19
- 238000012545 processing Methods 0.000 claims description 17
- 238000000926 separation method Methods 0.000 claims description 16
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 claims description 10
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 10
- 239000007995 HEPES buffer Substances 0.000 claims description 10
- 239000012980 RPMI-1640 medium Substances 0.000 claims description 10
- 239000012894 fetal calf serum Substances 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 229940054269 sodium pyruvate Drugs 0.000 claims description 10
- 229960005322 streptomycin Drugs 0.000 claims description 10
- 210000000496 pancreas Anatomy 0.000 claims description 9
- 230000006378 damage Effects 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 6
- 102100036790 Tubulin beta-3 chain Human genes 0.000 claims description 5
- 239000011159 matrix material Substances 0.000 claims description 4
- 238000003892 spreading Methods 0.000 claims description 4
- 101000878595 Arabidopsis thaliana Squalene synthase 1 Proteins 0.000 claims description 3
- 101000713575 Homo sapiens Tubulin beta-3 chain Proteins 0.000 claims description 3
- 210000000170 cell membrane Anatomy 0.000 abstract description 5
- 210000002950 fibroblast Anatomy 0.000 abstract description 4
- 230000005779 cell damage Effects 0.000 abstract description 3
- 208000037887 cell injury Diseases 0.000 abstract description 3
- 230000001276 controlling effect Effects 0.000 abstract description 3
- 230000001066 destructive effect Effects 0.000 abstract description 3
- 230000034659 glycolysis Effects 0.000 abstract description 3
- 230000005764 inhibitory process Effects 0.000 abstract description 3
- 230000001105 regulatory effect Effects 0.000 abstract description 3
- 230000000052 comparative effect Effects 0.000 description 9
- 102000029816 Collagenase Human genes 0.000 description 6
- 108060005980 Collagenase Proteins 0.000 description 6
- 229960002424 collagenase Drugs 0.000 description 6
- 210000004907 gland Anatomy 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- ZXKIFUWANBZSKF-UHFFFAOYSA-N [I].CC(O)=O Chemical compound [I].CC(O)=O ZXKIFUWANBZSKF-UHFFFAOYSA-N 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000000432 density-gradient centrifugation Methods 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 108010081750 Reticulin Proteins 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 210000003890 endocrine cell Anatomy 0.000 description 1
- 210000003020 exocrine pancreas Anatomy 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000000277 pancreatic duct Anatomy 0.000 description 1
- 238000011403 purification operation Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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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
- C12N5/0676—Pancreatic cells
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/05—Inorganic components
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Abstract
The invention discloses a method for treating hybrid cells on the surface of in vitro culture of pancreatic islets, which is to add iodoacetic acid with proper concentration into a pancreatic culture medium for culturing pancreatic islets and culture for a period of time. The invention utilizes the extremely strong destructive effect of the iodoacetic acid on cell membranes and the inhibition capability of the iodoacetic acid on the glycolysis of cells, and treats the hybrid cells (especially fibroblasts) on the surfaces of the pancreatic islets by regulating the concentration of the iodoacetic acid in the culture medium and controlling the treatment time, thereby reducing the subsequent experimental steps and reducing the cell damage.
Description
Technical Field
The invention relates to the technical field of cell culture, in particular to a method for treating hybrid cells on the surface of in vitro culture of pancreatic islets.
Background
Pancreatic islets refer to the mass of endocrine cells distributed between the acini of the exocrine pancreas. The surface of the islets is coated with a coating consisting of a thin layer of reticular fibers, but the coating of different species is not necessarily complete. Islets can be obtained by collagenase thermal digestion of the pancreas combined with density gradient centrifugation purification procedures. The pancreatic islet purification method comprises the steps of pouring collagenase solution with a proper concentration into a pancreatic gland through a pancreatic duct, picking the poured pancreatic gland, soaking the pancreatic gland into the collagenase solution, incubating the pancreatic gland in water bath at 37 ℃ for a certain time to enable the pancreatic gland to be digested into fine sand by the collagenase, stopping the digestion effect of the collagenase, primarily purifying the pancreatic islets by a density gradient centrifugation method, and picking the pancreatic islets by hands to perform secondary purification to obtain the pancreatic islets with extremely high purity. In order to obtain high-activity intact islets, collagenase is not suitable for a long time, so the purified islets generally contain a small amount of hybrid cells on the surface, including fibroblasts, acinar cells, ductal cells and the like.
The fibroblast and other hybrid cells can grow faster than the islet (cell) adherent growth, the excessively grown hybrid cells can obviously influence the adherent and growth states of the islet in vitro culture, and the development of islet research work is not facilitated. Therefore, how to effectively inhibit the growth of the hybrid cells (especially fibroblasts) is very important for the in vitro culture of the pancreatic islets. The proportion of the hybrid cells can be well reduced by multiple times of plate rotating culture, but the operation is time-consuming and labor-consuming, is not suitable for large-scale in-vitro culture of the pancreatic islets, and the intensive operation can also cause great adverse effect on the growth state of the pancreatic islets.
Accordingly, the prior art is deficient and needs improvement.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: provides a method for processing hybrid cells on the surface of in vitro culture of pancreatic islets, which removes the influence of the hybrid cells on the pancreatic islets.
The technical scheme of the invention is as follows: provides a method for processing hybrid cells on the surface of in vitro culture of pancreatic islets, which comprises the following steps: the following steps;
s1: spreading laminin in culture container to ensure homogeneous distribution of laminin solution on surface, and placing in CO2And (5) incubating the incubator.
S2: and selecting the islets to be cultured after separation and purification, and paving the islets on the incubated laminin.
S3: adding islet culture medium containing 0.5ng/ml-1 μ g/ml iodoacetic acid into culture container, and culturing for 0.5-120 h.
S4: after the culture is finished, the islet culture medium containing 0.5ng/ml-1 mug/ml iodoacetic acid is removed, and pure islets are obtained.
The iodine acetic acid can destroy cell membranes of the hybrid cells and the cells in the pancreatic islets, but the pancreatic islets are clustered cell clusters, so that the concentration of the iodine acetic acid and the culture time are adjusted, the cells on the surfaces of the pancreatic islets are also destroyed after the hybrid cells are destroyed, most of the cells in the pancreatic islets still survive and still serve as the pancreatic islets, the purity of the pancreatic islets is improved, and the subsequent operation of the pancreatic islets is facilitated.
The islet culture medium is prepared by adding the following components into an RPMI 1640 culture medium: 1.2g/L glucose, 10mmol/L HEPES, 10mg/L sodium pyruvate, 10% v/v fetal calf serum, 1% v/v penicillin-streptomycin; RPMI 1640 medium is commercially available, e.g., from sigma, USA under the designation R1383; the above glucose is commercially available, for example, from sigma, usa under the designation G7021; the HEPES described above is commercially available, e.g., from sigma, USA under the designation V900477; the sodium pyruvate is commercially available, for example, from sigma, usa under the designation P5280; the fetal calf serum is commercially available, such as from BIOLOGICAL INDUSTRIES, Israel, under the reference REF 03-031-1B; the above-mentioned penicillin-streptomycin is commercially available, for example, from BIOLOGICAL INDUSTRIES, Israel under the cat number 04-001-1 ACS.
In step S1, the laminin matrix cannot dry out.
The islets separated and purified in step S2 are cell masses obtained by digesting pancreas and purifying, and before selection, CO is added2Culturing overnight in an incubator to regulate and recover the damage of the islets during separation and purification.
In step S3, the concentration of iodoacetic acid is 1ng/ml-100ng/ml, and the culture time is 5h-72 h.
In step S4, the islet medium containing iodoacetic acid is removed and replaced with an islet medium for culture.
The other technical scheme of the invention is as follows: provides a method for processing hybrid cells on the surface of in vitro culture of pancreatic islets, which is characterized by comprising the following steps;
SS 1: the culture vessel was subjected to TC treatment for future use.
SS 2: and selecting the islets to be cultured after separation and purification, and paving the islets in a culture container for TC treatment.
SS 3: islet culture medium containing 0.5ng/ml-1 μ g/ml iodoacetic acid was added to the culture vessel in step SS2 and cultured for 0.5h-120 h.
SS 4: after the culture is finished, the islet culture medium containing 0.5ng/ml-1 mug/ml iodoacetic acid is removed, and pure islets are obtained.
Like the previous scheme, the iodoacetic acid can destroy cell membranes of the hybrid cells and the cells in the islets, but since the islets are clustered together, the concentration of the iodoacetic acid and the culture time are adjusted, so that the cells on the surfaces of the islets are also destroyed after the hybrid cells are destroyed, but most of the cells in the islets still survive and still serve as the islets, the purity of the islets is improved, and the subsequent operation of the islets is facilitated.
The islet culture medium is prepared by adding the following components into an RPMI 1640 culture medium: 1.2g/L glucose, 10mmol/L HEPES, 10mg/L sodium pyruvate, 10% v/v fetal calf serum, 1% v/v penicillin-streptomycin; RPMI 1640 medium is commercially available, e.g., from sigma, USA under the designation R1383; the above glucose is commercially available, for example, from sigma, usa under the designation G7021; the HEPES described above is commercially available, e.g., from sigma, USA under the designation V900477; the sodium pyruvate is commercially available, for example, from sigma, usa under the designation P5280; the fetal calf serum is commercially available, such as from BIOLOGICAL INDUSTRIES, Israel, under the reference REF 03-031-1B; the above-mentioned penicillin-streptomycin is commercially available, for example, from BIOLOGICAL INDUSTRIES, Israel under the cat number 04-001-1 ACS.
The separated and purified islets of Langerhans in step SS2 are cell masses obtained by digesting pancreas and purifying, and before selecting, CO is added2Culturing overnight in an incubator to regulate and recover the damage of the islets during separation and purification.
In step SS3, the concentration of iodoacetic acid is 1ng/ml-100ng/ml, and the culture time is 5h-72 h.
In step SS4, the culture medium of islets was changed to islet culture medium after removal of the iodine-containing acetic acid.
By adopting the scheme, the invention provides the method for treating the hybrid cells on the surface of the islet in vitro culture, which utilizes the extremely strong destructive effect of the iodoacetic acid on cell membranes and the inhibition capability of the iodoacetic acid on the glycolysis of cells, and treats the hybrid cells on the surface of the islet by regulating the concentration of the iodoacetic acid in the culture medium and controlling the treatment time, so that the subsequent experimental steps are reduced, and the cell damage is reduced.
Drawings
FIG. 1 is a photograph of islets cultured in example 1 of the present invention;
FIG. 2 is a photograph of islets cultured in comparative example 1;
FIG. 3 is a photograph of islets cultured in example 2 of the present invention;
FIG. 4 is a photograph of islets cultured in comparative example 2.
Detailed Description
The invention is described in detail below with reference to the figures and the specific embodiments.
Example 1
The embodiment provides a method for processing hybrid cells on the surface of in vitro culture of pancreatic islets, which comprises the following steps: the following steps;
s1: laminin was spread in culture vessels to ensure that the laminin solution was evenly distributed on the surface, and then incubated in a CO2 incubator. In this step, the laminin matrix cannot dry out.
S2: and selecting the islets to be cultured after separation and purification, and paving the islets on the incubated laminin.
S3: islet medium containing 5ng/ml iodoacetic acid was added to the culture vessel and incubated for 20 h.
S4: after the culture is finished, removing the islet culture medium containing 5ng/ml iodoacetic acid to obtain pure islets, and adding the islet culture medium containing no iodoacetic acid into the pure islets for culture.
The islet culture medium is prepared by adding the following components into an RPMI 1640 culture medium: 1.2g/L glucose, 10mmol/L HEPES, 10mg/L sodium pyruvate, 10% v/v fetal calf serum, 1% v/v penicillin-streptomycin; RPMI 1640 medium is commercially available, e.g., from sigma, USA under the designation R1383; the above glucose is commercially available, for example, from sigma, usa under the designation G7021; the HEPES described above is commercially available, e.g., from sigma, USA under the designation V900477; the sodium pyruvate is commercially available, for example, from sigma, usa under the designation P5280; the fetal calf serum is commercially available, such as from BIOLOGICAL INDUSTRIES, Israel, under the reference REF 03-031-1B; the above-mentioned penicillin-streptomycin is commercially available, for example, from BIOLOGICAL INDUSTRIES, Israel under the cat number 04-001-1 ACS.
The islets separated and purified in step S2 are cell clusters obtained by digesting the pancreas and performing purification operations, and before selection, the islets are cultured overnight in a CO2 incubator, so that the damage of the islets during separation and purification can be adjusted and recovered.
Comparative example 1
S11: spreading laminin in culture container to ensure homogeneous distribution of laminin solution on surface, and placing in CO2And (5) incubating the incubator. In this step, the laminin matrix cannot dry out.
S12: and selecting the islets to be cultured after separation and purification, and paving the islets on the incubated laminin. The separated and purified islets are cell masses obtained by digesting pancreas and purifying, and before selection, the islets are cultured in a CO2 incubator overnight, so that the damage of the islets during separation and purification is adjusted and recovered.
S13: culturing for 20 h.
Referring to fig. 1 and 2, fig. 1 is a photograph of pancreatic islets obtained after the culture of example 1, fig. 2 is a photograph of pancreatic islets obtained after the culture of comparative example 1, and scales in fig. 1 and 2 are both 100 μm, and as can be seen from fig. 1 and 2, fig. 1 contains few hybrid cells, and fig. 2 contains many hybrid cells, it can be seen from example 1 and comparative example 1 that iodoacetic acid can effectively remove the hybrid cells of pancreatic islets, thereby facilitating the subsequent operation.
Example 2
The embodiment provides a method for processing hybrid cells on the surface of in vitro culture of pancreatic islets, which is characterized by comprising the following steps;
SS 1: the culture vessel was subjected to TC treatment for future use.
SS 2: and selecting the islets to be cultured after separation and purification, and paving the islets in a culture container for TC treatment.
SS 3: islet medium containing 5ng/ml iodoacetic acid was added to the culture vessel in step SS2 and incubated for 30 h.
SS 4: after the culture is finished, removing the islet culture medium containing 5ng/ml iodoacetic acid to obtain pure islets, and adding the islet culture medium containing no iodoacetic acid into the pure islets for culture.
The islet culture medium is prepared by adding the following components into an RPMI 1640 culture medium: 1.2g/L glucose, 10mmol/L HEPES, 10mg/L sodium pyruvate, 10% v/v fetal calf serum, 1% v/v penicillin-streptomycin; RPMI 1640 medium is commercially available, e.g., from sigma, USA under the designation R1383; the above glucose is commercially available, for example, from sigma, usa under the designation G7021; the HEPES described above is commercially available, e.g., from sigma, USA under the designation V900477; the sodium pyruvate is commercially available, for example, from sigma, usa under the designation P5280; the fetal calf serum is commercially available, such as from BIOLOGICAL INDUSTRIES, Israel, under the reference REF 03-031-1B; the above-mentioned penicillin-streptomycin is commercially available, for example, from BIOLOGICAL INDUSTRIES, Israel under the cat number 04-001-1 ACS.
The separated and purified islets of Langerhans in step SS2 are cell masses obtained by digesting pancreas and purifying, and before selecting, CO is added2Culturing overnight in an incubator to regulate and recover the damage of the islets during separation and purification.
Comparative example 2
SS 11: the culture vessel was subjected to TC treatment for future use.
SS 12: and selecting the islets to be cultured after separation and purification, and paving the islets in a culture container for TC treatment. The separated and purified islets are cell masses obtained by digesting pancreas and purifying, and before selection, the islets are cultured in a CO2 incubator overnight, so that the damage of the islets during separation and purification is adjusted and recovered.
SS 13: islet medium was added to the culture vessel in step SS12 and cultured for 30 h.
Referring to fig. 3 and 4, fig. 3 is a photograph of islets obtained after the culture of example 2, fig. 4 is a photograph of islets obtained after the culture of comparative example 2, and scales in fig. 3 and 4 are both 100 μm, and as can be seen from fig. 3 and 4, fig. 3 contains few hybrid cells, and fig. 4 contains many hybrid cells, it can be seen from example 2 and comparative example 2 that iodoacetic acid can effectively remove the hybrid cells of islets, thereby facilitating the subsequent operation.
Therefore, as can be seen from examples 1 and 2, and comparative examples 1 and 2, the addition of iodoacetic acid can effectively remove the islet hybrid cells, and facilitate subsequent operations.
In conclusion, the invention provides a method for processing hybrid cells on the surface of in vitro culture of pancreatic islets, which utilizes the extremely strong destructive effect of iodoacetic acid on cell membranes and the inhibition capability of the iodoacetic acid on the glycolysis of cells, and processes the hybrid cells on the surfaces of the pancreatic islets by regulating the concentration of the iodoacetic acid in a culture medium and controlling the processing time, thereby reducing the subsequent experimental steps and reducing the cell damage.
The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A method for processing hybrid cells on the surface of in vitro culture of pancreatic islets is characterized by comprising the following steps:
s1: spreading laminin in culture container to ensure homogeneous distribution of laminin solution on surface, and placing in CO2Incubating in an incubator;
s2: selecting the islets to be cultured after separation and purification and spreading the islets on the incubated laminin;
s3: adding islet culture medium containing 0.5ng/ml-1 μ g/ml iodoacetic acid into culture container, and culturing for 0.5-120 h;
s4: after the culture is finished, the islet culture medium containing 0.5ng/ml-1 mug/ml iodoacetic acid is removed, and pure islets are obtained.
2. The method for processing the hybrid cells on the surface of the islet in vitro culture according to claim 1, wherein the islet culture medium is RPMI 1640 medium added with the following components: 1.2g/L glucose, 10mmol/L HEPES, 10mg/L sodium pyruvate, 10% v/v fetal calf serum, 1% v/v penicillin-streptomycin.
3. The method for processing hybrid cells on the surface of in vitro culture of pancreatic islets according to claim 1, wherein in step S1, the laminin matrix cannot be dried; the islets of Langerhans in step S2 are cell clusters obtained by digesting the pancreas and purifying, and are first treated with CO before selection2Culturing overnight in an incubator to regulate and recover the damage of the islets during separation and purification.
4. The method for processing hybrid cells on the surface of in vitro culture of pancreatic islets according to claim 1, wherein the concentration of iodoacetic acid is 1ng/ml to 100ng/ml and the culture time is 5h to 72h in step S3.
5. The method for processing hybrid cells on the surface of in vitro culture of pancreatic islets according to claim 1, wherein the pancreatic islet culture medium containing iodoacetic acid is removed and replaced with the pancreatic islet culture medium for culture in step S4.
6. A method for processing hybrid cells on the surface of in vitro culture of pancreatic islets is characterized by comprising the following steps:
SS 1: performing TC treatment on the culture container for later use;
SS 2: selecting the islets to be cultured after separation and purification and paving the islets in a culture container for TC treatment;
SS 3: adding islet culture medium containing 0.5ng/ml-1 μ g/ml iodoacetic acid into the culture container in step SS2, and culturing for 0.5-120 h;
SS 4: after the culture is finished, the islet culture medium containing 0.5ng/ml-1 mug/ml iodoacetic acid is removed, and pure islets are obtained.
7. The method for processing hybrid cells on the surface of in vitro culture of pancreatic islets according to claim 6, wherein the pancreatic islet culture medium is RPMI 1640 medium supplemented with the following components: 1.2g/L glucose, 10mmol/L HEPES, 10mg/L sodium pyruvate, 10% v/v fetal calf serum, 1% v/v penicillin-streptomycin.
8. The method for processing hybrid cells on the surface of islet in vitro culture according to claim 6, wherein the isolated and purified islets in step SS2 are cell clusters obtained by digesting pancreas and purifying, and before selection, CO is firstly added2Culturing overnight in an incubator to regulate and recover the damage of the islets during separation and purification.
9. The method for processing hybrid cells on the surface of islet in vitro culture according to claim 6, wherein in step SS3, the concentration of iodoacetic acid is 1ng/ml-100ng/ml, and the culture time is 5h-72 h.
10. The method for processing hybrid cells on the surface of islet in vitro culture according to claim 6, wherein in step SS4, the islet culture medium containing iodoacetic acid is removed and replaced with the islet culture medium for culture.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5681587A (en) * | 1995-10-06 | 1997-10-28 | Desmos, Inc. | Growth of adult pancreatic islet cells |
| WO2000078929A1 (en) * | 1999-06-23 | 2000-12-28 | Joslin Diabetes Center, Inc. | Methods of making pancreatic islet cells |
| US20020072115A1 (en) * | 2000-02-18 | 2002-06-13 | The Walter And Eliza Hall Institute Of Medical Research | Pancreatic islet cell growth factors |
| WO2019098944A1 (en) * | 2017-11-15 | 2019-05-23 | National University Of Singapore | Systems for expanding pancreatic islets and transplantation thereof |
-
2021
- 2021-04-27 CN CN202110460519.8A patent/CN113308433A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5681587A (en) * | 1995-10-06 | 1997-10-28 | Desmos, Inc. | Growth of adult pancreatic islet cells |
| WO2000078929A1 (en) * | 1999-06-23 | 2000-12-28 | Joslin Diabetes Center, Inc. | Methods of making pancreatic islet cells |
| US20020072115A1 (en) * | 2000-02-18 | 2002-06-13 | The Walter And Eliza Hall Institute Of Medical Research | Pancreatic islet cell growth factors |
| WO2019098944A1 (en) * | 2017-11-15 | 2019-05-23 | National University Of Singapore | Systems for expanding pancreatic islets and transplantation thereof |
Non-Patent Citations (1)
| Title |
|---|
| 张晓春: "降糖孜雅比提片（HZT）对体外培养胰岛细胞的影响", 新疆医科大学硕士论文, no. 09, 15 September 2008 (2008-09-15), pages 5 * |
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