CN109628385A - A kind of human oocyte In-vitro maturation liquid and preparation method thereof and cultural method - Google Patents
A kind of human oocyte In-vitro maturation liquid and preparation method thereof and cultural method Download PDFInfo
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- CN109628385A CN109628385A CN201910049864.5A CN201910049864A CN109628385A CN 109628385 A CN109628385 A CN 109628385A CN 201910049864 A CN201910049864 A CN 201910049864A CN 109628385 A CN109628385 A CN 109628385A
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- 210000000287 oocyte Anatomy 0.000 title claims abstract description 78
- 230000035800 maturation Effects 0.000 title claims abstract description 48
- 238000000338 in vitro Methods 0.000 title claims abstract description 39
- 239000007788 liquid Substances 0.000 title claims abstract description 36
- 238000000034 method Methods 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 claims abstract description 26
- 229940054269 sodium pyruvate Drugs 0.000 claims abstract description 13
- 101100468640 Danio rerio rhcgl2 gene Proteins 0.000 claims abstract description 12
- 101150053759 rhcg gene Proteins 0.000 claims abstract description 12
- 238000004113 cell culture Methods 0.000 claims abstract description 11
- 102000002322 Egg Proteins Human genes 0.000 claims description 35
- 108010000912 Egg Proteins Proteins 0.000 claims description 35
- 210000004681 ovum Anatomy 0.000 claims description 35
- 210000000130 stem cell Anatomy 0.000 claims description 30
- 238000006243 chemical reaction Methods 0.000 claims description 20
- 210000004508 polar body Anatomy 0.000 claims description 10
- 210000001771 cumulus cell Anatomy 0.000 claims description 7
- 238000001976 enzyme digestion Methods 0.000 claims description 7
- 229920002472 Starch Polymers 0.000 claims description 5
- 210000001733 follicular fluid Anatomy 0.000 claims description 5
- 239000011265 semifinished product Substances 0.000 claims description 5
- 235000019698 starch Nutrition 0.000 claims description 5
- 239000008107 starch Substances 0.000 claims description 5
- 230000001256 tonic effect Effects 0.000 claims description 5
- 239000001963 growth medium Substances 0.000 abstract description 11
- 210000002459 blastocyst Anatomy 0.000 abstract description 7
- 238000003776 cleavage reaction Methods 0.000 abstract description 5
- 230000007017 scission Effects 0.000 abstract description 5
- 210000004027 cell Anatomy 0.000 abstract description 4
- 210000001161 mammalian embryo Anatomy 0.000 abstract description 4
- 230000021121 meiosis Effects 0.000 abstract description 4
- 230000011748 cell maturation Effects 0.000 abstract description 3
- 230000001086 cytosolic effect Effects 0.000 abstract description 3
- 230000000694 effects Effects 0.000 abstract description 3
- 231100000331 toxic Toxicity 0.000 abstract description 3
- 230000002588 toxic effect Effects 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 2
- 230000015572 biosynthetic process Effects 0.000 description 4
- 230000004927 fusion Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000011177 media preparation Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 210000002380 oogonia Anatomy 0.000 description 2
- 102100038343 Ammonium transporter Rh type C Human genes 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 101000666627 Homo sapiens Ammonium transporter Rh type C Proteins 0.000 description 1
- 206010033266 Ovarian Hyperstimulation Syndrome Diseases 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000034004 oogenesis Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 210000000801 secondary oocyte Anatomy 0.000 description 1
Classifications
-
- 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
- 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
- C12N5/0608—Germ cells
- C12N5/0609—Oocytes, oogonia
-
- 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
- C12N2500/30—Organic components
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/31—Pituitary sex hormones, e.g. follicle-stimulating hormone [FSH], luteinising hormone [LH]; Chorionic gonadotropins
Abstract
The invention discloses a kind of human oocyte In-vitro maturation liquid and preparation method thereof and cultural methods, it is related to medicine technology field, it includes following components in terms of mass fraction that it, which is formulated: it includes following components in terms of mass fraction that it, which is formulated: 70-95 parts of 75IU/L rFSH, 60-70 parts of 150IU/L rHCG, 50-55 parts of 20%SPS, 30-40 parts of 2mg/L EGF, 150-200 parts of 20-25 parts of 25mmol/L Sodium Pyruvate and elementary cell culture solution Qiunn ' s 1029.The people's oocyte in vitro maturation culture solution and preparation method thereof and cultural method, the cost of purchase commercialization In-vitro maturation base can be effectively reduced, the culture medium is basic culture medium with people blastocyst culture liquid Qiunn ' s 1029, the culture medium safe without toxic side effect, it can promote the Oocyte Meiosis in immature oocyte especially three-level cumulus oocytes complesxes, improve the ectogenesis ability of such cell, promote Cytoplasmic maturation, cleavage rates and blastocyst rate after improving cell maturation, further increase embryo quality.
Description
Technical field
The present invention relates to medicine technology field, specially a kind of human oocyte In-vitro maturation liquid and preparation method thereof
And cultural method.
Background technique
Egg mother cell refer in Process of oogenesis carry out meiosis oogonium, be divided into first oocyte,
Secondary oocyte and mature egg mother cell, they are to generate after oogonium differentiation is divided with DNA replication dna, for the first time respectively
The product of meiosis and second meiotic division.
Ovum mother not only can be improved as the important component as assisted reproductive technology in egg mother cell external mature technology
Cell survives efficiency, reduces the generation of ovarian hyperstimulation syndrome, also for give ovum female reproduction try hard to keep deposit provide it is more
Selection, but the higher cost of the preparation method purchase commercialization In-vitro maturation base of existing culture solution, human oocyte body
Outer maturing rate is low, and culture medium preparation method and cultural method are complicated, for this purpose, we provide a kind of human oocyte maturation in vitro
Culture solution and preparation method thereof and cultural method solve the problems, such as this.
Summary of the invention
The object of the invention is in order to make up for the deficiencies of the prior art, provide a kind of human oocyte maturation in vitro training
Nutrient solution and preparation method thereof and cultural method, it, which has, improves human oocyte maturation in vitro rate, simplified culture base preparation method
And cultural method, the advantages of cost of In-vitro maturation base is commercialized in purchase is reduced, solves human oocyte maturation in vitro
The problem of rate is low, and culture medium preparation method and cultural method are complicated, the higher cost of purchase commercialization In-vitro maturation base.
The present invention is in order to solve the above technical problems, provide the following technical solutions: a kind of human oocyte In-vitro maturation
Liquid, formula include following components in terms of mass fraction: 70-95 parts of 75IU/L rFSH, 60-70 parts of 150IU/L rHCG,
50-55 parts of 20%SPS, 30-40 parts of 2mg/L EGF, 20-25 parts of 25mmol/L Sodium Pyruvate and elementary cell culture solution
Qiunn ' s 1029 150-200 part.
Further, preparation method such as following steps:
Elementary cell culture solution Qiunn ' s 1029 is put into inside reaction kettle by S1 first, and be added 75IU/L rFSH and
150IU/L rHCG, starts reaction kettle later, is sufficiently mixed it, and place 30 minutes merges it sufficiently later；
20%SPS, 2mg/L EGF and 25mmol/L Sodium Pyruvate are successively sequentially added inside reaction kettle, and started simultaneously by S2
Reaction kettle, temperature are maintained at 5 DEG C, and mixing mixes well it in 30 minutes；
Semi-finished product obtained in step S2 are put into inside medical refrigerator by S3, and internal temperature of refrigerator is maintained at 2-8 DEG C, has prepared
At.
Further, cultural method the following steps are included:
S1, takes ovum proxima luce (prox. luc) in patient, and human oocyte In-vitro maturation liquid is put into constant incubator internal balance 16-18
Hour；
S2, patient take the daily sliding scale identification of ovum and pick up the immature oocyte in liquor folliculi, the defeated ovum of conventional H EPE ' s people
It is observed under inverted microscope after pipe liquid rinses, the MI phase egg mother cell and ooecium that first polar body is not discharged starch interior still visible hair tonic
The GV phase egg mother cell of bubble is immature oocyte, reuses human oocyte In-vitro maturation liquid and is rinsed, with
People's immature oocyte is put into human oocyte In-vitro maturation liquid 1mL afterwards, 5%O in constant incubator2、5%CO2、
Culture 26-28 hours is carried out under the conditions of 37 DEG C；
S3 takes out the human oocyte that culture is completed in step S2, and shelling ovum needle after enzymic digestion will be around human oocyte body
Cumulus cell strips；
The step S5 human oocyte body for stripping completion is finally placed under inverted microscope, further looks at and confirm ovum by S4
First polar body is discharged with egg mother cell in the mature condition of mother cell, becomes the mark that MII phase egg mother cell is maturation of ovum.
Further, described to be stripped the cumulus cell around human oocyte body by shelling ovum needle after enzymic digestion.
Further, the insulating box internal temperature should remain 37 DEG C, CO2Concentration is maintained at 5%, O2Concentration is maintained at
5%。。
Compared with prior art, a kind of human oocyte In-vitro maturation liquid and preparation method thereof and cultural method tool
It is standby following the utility model has the advantages that
1, the present invention can be effectively reduced the cost of purchase commercialization In-vitro maturation base, save money by the preparation method
Source is conducive to improve production capacity and production efficiency.
2, the present invention is basic culture medium, interior addition with people blastocyst culture liquid Qiunn ' s 1029 by the culture medium
75IU/L rFSH, 150IU/L rHCG, 20%SPS, 2mg/L EGF, 25mmol/L Sodium Pyruvate and be made, the culture medium safety
It has no toxic side effect, the egg mother cell in immature oocyte especially three-level cumulus oocytes complesxes can be promoted to subtract
Number division, improves the ectogenesis ability of such cell, promotes Cytoplasmic maturation, the cleavage rates and blastaea after improving cell maturation
Formation rate further increases embryo quality.
3, the Nuclear maturity rate for obtaining human oocyte that the present invention is made by the cultural method is higher, can achieve 90%, ovum
Mother cell matter is mature more preferable, and the cleavage rates and Blastocyst formation rate after maturation are higher, further increases embryo quality.
Specific embodiment
Based on the embodiments of the present invention, those of ordinary skill in the art are obtained without making creative work
The every other embodiment obtained, shall fall within the protection scope of the present invention.
Embodiment one
A kind of human oocyte In-vitro maturation liquid, it is characterised in that: 70 parts of 75IU/L rFSH, 150IU/L rHCG 60
Part, 50 parts of 20%SPS, 30 parts of 2mg/L EGF, 20 parts of 25mmol/L Sodium Pyruvate and elementary cell culture solution Qiunn ' s
1029 150 parts.
Preparation method such as following steps:
1,029 150 parts of s of elementary cell culture solution Qiunn ' are put into inside reaction kettle first, and 75IU/L rFSH are added by S1
70 parts and 60 parts of 150IU/L rHCG, start reaction kettle later, be sufficiently mixed it, place 30 minutes fills it later
Divide fusion；
S2 successively sequentially adds 50 parts of 20%SPS, EGF30 parts of 2mg/L and 20 parts of 25mmol/L Sodium Pyruvate in reaction kettle
Portion, and start reaction kettle simultaneously, temperature is maintained at 5 DEG C, and mixing mixes well it in 30 minutes；
Semi-finished product obtained in step S2 are put into inside medical refrigerator by S3, and internal temperature of refrigerator is maintained at 2-8 DEG C, has prepared
At.
Its cultural method the following steps are included:
S1, takes ovum proxima luce (prox. luc) in patient, and human oocyte In-vitro maturation liquid is put into constant incubator internal balance 16-18
Hour；
S2, patient take the daily sliding scale identification of ovum and pick up the immature oocyte in liquor folliculi, the defeated ovum of conventional H EPE ' s people
It is observed under inverted microscope after pipe liquid rinses, the MI phase egg mother cell and ooecium that first polar body is not discharged starch interior still visible hair tonic
The GV phase egg mother cell of bubble is immature oocyte, reuses human oocyte In-vitro maturation liquid and is rinsed, with
People's immature oocyte is put into human oocyte In-vitro maturation liquid 1mL afterwards, 5%O in constant incubator2、5%CO2、
Culture 26-28 hours is carried out under the conditions of 37 DEG C；
S3 takes out the human oocyte that culture is completed in step S2, and shelling ovum needle after enzymic digestion will be around human oocyte body
Cumulus cell strips；
The step S5 human oocyte body for stripping completion is finally placed under inverted microscope, further looks at and confirm ovum by S4
First polar body is discharged with egg mother cell in the mature condition of mother cell, becomes the mark that MII phase egg mother cell is maturation of ovum.
Embodiment two
A kind of human oocyte In-vitro maturation liquid, it is characterised in that: 80 parts of 75IU/L rFSH, 150IU/L rHCG 65
Part, 53 parts of 20%SPS, 30 parts of 2mg/L EGF, 23 parts of 25mmol/L Sodium Pyruvate and elementary cell culture solution Qiunn ' s
1029 170 parts.
Preparation method such as following steps:
1,029 170 parts of s of elementary cell culture solution Qiunn ' are put into inside reaction kettle first, and 75IU/L rFSH are added by S1
80 parts and 65 parts of 150IU/L rHCG, start reaction kettle later, be sufficiently mixed it, place 30 minutes fills it later
Divide fusion；
S2 successively sequentially adds 53 parts of 20%SPS, EGF30 parts of 2mg/L and 23 parts of 25mmol/L Sodium Pyruvate in reaction kettle
Portion, and start reaction kettle simultaneously, temperature is maintained at 5 DEG C, and mixing mixes well it in 30 minutes；
Semi-finished product obtained in step S2 are put into inside medical refrigerator by S3, and internal temperature of refrigerator is maintained at 2-8 DEG C, has prepared
At.
Its cultural method the following steps are included:
S1, takes ovum proxima luce (prox. luc) in patient, and human oocyte In-vitro maturation liquid is put into constant incubator internal balance 16-18
Hour；
S2, patient take the daily sliding scale identification of ovum and pick up the immature oocyte in liquor folliculi, the defeated ovum of conventional H EPE ' s people
It is observed under inverted microscope after pipe liquid rinses, the MI phase egg mother cell and ooecium that first polar body is not discharged starch interior still visible hair tonic
The GV phase egg mother cell of bubble is immature oocyte, reuses human oocyte In-vitro maturation liquid and is rinsed, with
People's immature oocyte is put into human oocyte In-vitro maturation liquid 1mL afterwards, 5%O in constant incubator2、5%CO2、
Culture 26-28 hours is carried out under the conditions of 37 DEG C；
S3 takes out the human oocyte that culture is completed in step S2, and shelling ovum needle after enzymic digestion will be around human oocyte body
Cumulus cell strips；
The step S5 human oocyte body for stripping completion is finally placed under inverted microscope, further looks at and confirm ovum by S4
First polar body is discharged with egg mother cell in the mature condition of mother cell, becomes the mark that MII phase egg mother cell is maturation of ovum.
Embodiment three
A kind of human oocyte In-vitro maturation liquid, it is characterised in that: 95 parts of 75IU/L rFSH, 150IU/L rHCG 70
Part, 55 parts of 20%SPS, 40 parts of 2mg/L EGF, 25 parts of 25mmol/L Sodium Pyruvate and elementary cell culture solution Qiunn ' s
1029 200 parts.
Preparation method such as following steps:
1,029 200 parts of s of elementary cell culture solution Qiunn ' are put into inside reaction kettle first, and 75IU/L rFSH are added by S1
95 parts and 70 parts of 150IU/L rHCG, start reaction kettle later, be sufficiently mixed it, place 30 minutes fills it later
Divide fusion；
S2 successively sequentially adds 55 parts of 20%SPS, EGF40 parts of 2mg/L and 25 parts of 25mmol/L Sodium Pyruvate in reaction kettle
Portion, and start reaction kettle simultaneously, temperature is maintained at 5 DEG C, and mixing mixes well it in 30 minutes；
Semi-finished product obtained in step S2 are put into inside medical refrigerator by S3, and internal temperature of refrigerator is maintained at 2-8 DEG C, has prepared
At.
Its cultural method the following steps are included:
S1, takes ovum proxima luce (prox. luc) in patient, and human oocyte In-vitro maturation liquid is put into constant incubator internal balance 16-18
Hour；
S2, patient take the daily sliding scale identification of ovum and pick up the immature oocyte in liquor folliculi, the defeated ovum of conventional H EPE ' s people
It is observed under inverted microscope after pipe liquid rinses, the MI phase egg mother cell and ooecium that first polar body is not discharged starch interior still visible hair tonic
The GV phase egg mother cell of bubble is immature oocyte, reuses human oocyte In-vitro maturation liquid and is rinsed, with
People's immature oocyte is put into human oocyte In-vitro maturation liquid 1mL afterwards, 5%O in constant incubator2、5%CO2、
Culture 26-28 hours is carried out under the conditions of 37 DEG C；
S3 takes out the human oocyte that culture is completed in step S2, and shelling ovum needle after enzymic digestion will be around human oocyte body
Cumulus cell strips；
The step S5 human oocyte body for stripping completion is finally placed under inverted microscope, further looks at and confirm ovum by S4
First polar body is discharged with egg mother cell in the mature condition of mother cell, becomes the mark that MII phase egg mother cell is maturation of ovum.
It randomly selects the human oocyte body that culture is completed to be detected, testing result is as follows:
| Classification | Maturing rate | Cleavage rates | Blastocyst formation rate |
| Embodiment one | 93% | 92% | 91% |
| Embodiment two | 95% | 94% | 93% |
| Embodiment three | 97% | 97% | 93.5% |
The present invention is basic culture medium with people's blastocyst culture by the culture medium, interior to add 75IU/L rFSH, 150IU/L
RHCG, 20%SPS, 2mg/L EGF, 25mmol/L Sodium Pyruvate and be made, the culture medium safe without toxic side effect, can make not
Oocyte Meiosis in mature oocyte especially three-level cumulus oocytes complesxes, improves such cell
Ectogenesis ability promotes Cytoplasmic maturation, and cleavage rates and Blastocyst formation rate after improving cell maturation further increase embryo
Quality.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (5)
1. a kind of human oocyte In-vitro maturation liquid, it is characterised in that: it includes following components in terms of mass fraction that it, which is formulated:
70-95 parts of 75IU/L rFSH, 60-70 parts of 150IU/L rHCG, 20% 50-55 parts of SPS, 30-40 parts of 2mg/L EGF,
150-200 parts of 20-25 parts of 25mmol/L Sodium Pyruvate and elementary cell culture solution Qiunn ' s 1029.
2. a kind of preparation method of human oocyte In-vitro maturation liquid according to claim 1, it is characterised in that: its
Preparation method such as following steps:
Elementary cell culture solution Qiunn ' s 1029 is put into inside reaction kettle by S1 first, and be added 75IU/L rFSH and
150IU/L rHCG, starts reaction kettle later, is sufficiently mixed it, and place 30 minutes merges it sufficiently later；
20%SPS, 2mg/L EGF and 25mmol/L Sodium Pyruvate are successively sequentially added inside reaction kettle, and started simultaneously by S2
Reaction kettle, temperature are maintained at 5 DEG C, and mixing mixes well it in 30 minutes；
Semi-finished product obtained in step S2 are put into inside medical refrigerator by S3, and internal temperature of refrigerator is maintained at 2-8 DEG C, has prepared
At.
3. a kind of cultural method of human oocyte In-vitro maturation liquid according to claim 2, it is characterised in that: its
Cultural method the following steps are included:
S1, takes ovum proxima luce (prox. luc) in patient, and human oocyte In-vitro maturation liquid is put into constant incubator internal balance 16-18
Hour；
S2, patient take the daily sliding scale identification of ovum and pick up the immature oocyte in liquor folliculi, the defeated ovum of conventional H EPE ' s people
It is observed under inverted microscope after pipe liquid rinses, the MI phase egg mother cell and ooecium that first polar body is not discharged starch interior still visible hair tonic
The GV phase egg mother cell of bubble is immature oocyte, reuses human oocyte In-vitro maturation liquid and is rinsed, with
People's immature oocyte is put into human oocyte In-vitro maturation liquid 1mL afterwards, 5%O in constant incubator2、5%CO2、
Culture 26-28 hours is carried out under the conditions of 37 DEG C；
S3 takes out the human oocyte that culture is completed in step S2, and shelling ovum needle after enzymic digestion will be around human oocyte body
Cumulus cell strips；
The step S5 human oocyte body for stripping completion is finally placed under inverted microscope, further looks at and confirm ovum by S4
First polar body is discharged with egg mother cell in the mature condition of mother cell, becomes the mark that MII phase egg mother cell is maturation of ovum.
4. a kind of cultural method of human oocyte In-vitro maturation liquid according to claim 3, it is characterised in that: institute
It states by shelling the cumulus cell around ovum needle mechanical removal human oocyte body after enzymic digestion.
5. a kind of cultural method of human oocyte In-vitro maturation liquid according to claim 3, it is characterised in that: institute
Should be remained by stating insulating box internal temperature by 37 DEG C, CO2Concentration is maintained at 5%, O2Concentration is maintained at 5%.
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| CN112646771A (en) * | 2021-01-22 | 2021-04-13 | 艾尔斯（浙江）医学科技有限公司 | Cell line of human follicle cumulus cells or/and granulosa cells and preparation method thereof |
| CN112680406A (en) * | 2021-01-12 | 2021-04-20 | 艾尔斯（浙江）医学科技有限公司 | Fertility preservation method for egg production through caesarean section |
| CN114164168A (en) * | 2021-11-25 | 2022-03-11 | 苏州原一医疗科技有限公司 | In-vitro maturation culture solution and culture method for human oocyte-cumulus granular cell complex |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112680406A (en) * | 2021-01-12 | 2021-04-20 | 艾尔斯（浙江）医学科技有限公司 | Fertility preservation method for egg production through caesarean section |
| CN112646771A (en) * | 2021-01-22 | 2021-04-13 | 艾尔斯（浙江）医学科技有限公司 | Cell line of human follicle cumulus cells or/and granulosa cells and preparation method thereof |
| CN114164168A (en) * | 2021-11-25 | 2022-03-11 | 苏州原一医疗科技有限公司 | In-vitro maturation culture solution and culture method for human oocyte-cumulus granular cell complex |
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