WO1986007377A1 - Culture media for in vitro fertilization and embryo transfer - Google Patents
Culture media for in vitro fertilization and embryo transfer Download PDFInfo
- Publication number
- WO1986007377A1 WO1986007377A1 PCT/AU1986/000170 AU8600170W WO8607377A1 WO 1986007377 A1 WO1986007377 A1 WO 1986007377A1 AU 8600170 W AU8600170 W AU 8600170W WO 8607377 A1 WO8607377 A1 WO 8607377A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- sodium
- potassium
- culture medium
- ions
- chloride
- Prior art date
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Classifications
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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
- 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
- C12N2500/05—Inorganic 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/34—Sugars
Definitions
- the invention relates to culture media and more particularly to culture media for in vitro fertilization and embryo transfer.
- n vitro fertilization of human oocytes, cleavage of embryos and embryo transfer require that a culture medium be used to support the embryo for a period of up to three or four days during the various processes necessary for fertilization and early incubation before embryo transfer and reimplantatlon.
- the oocyte In the natural process of fertilization for a human oocyte the oocyte is supported within the mother within a fluid known as human tubal fluid andi t i s the object of the present invention to provide a culture medium as a synthetic human tubal fluid.
- the present invention we have devised a synthetic culture medium which is believed to approximate human tubal fluid but with desirable additional components and variations in the actual composition, including ratios of concentrations of sodium ions to potassium ions.
- the present invention may be said to reside in a culture medium for in vitro fertilization and embryo transfer comprising the following compounds in the following ranges of concentration;
- the ratio of sodium ion concentration to potassium ion concentration is in the range from 28 to 32.
- the ratio of concentrations of sodium ions to potassium ions is 29.3.
- the synthetic human tubal fluid may have the following composition
- the invention may be said to reside in a method of assisting with the in vitro fertilization of human oocytes including the step of handling the human oocytes in a culture medium, the culture medium being comprised of the compounds listed below in the range of compositions listed as follows;
- the ratio of sodium ion concentration to potassium ion concentration is in the range of from 28 to 32.
- the ratio concentration of sodium ions to potassium ions is 29.3.
- the method includes the step of handling the oocytes in a culture medium comprising;
- thei nvention may be said to reside in a culture medium for thei n vitro fertilization of human oocytesi ncluding sodium potassiumi ons wherein the ratio of sodium ions to potassium ions is i n the range of from 28 to 32.
- the ratio of sodium ions to potassium ions is 29.3.
- synthetic human tubal fluid culture medium is as given in Table 1 below (marked synthetic HTF).
- the medium may be prepared by using rainwater which has been distilled in glass six times.
- the bicarbonate-buffered medium is gassed for a minimum of five minutes with humidified 5% oxygen 5% carbon dioxide 90% nitrogen mixture and sterilized by passage through a 0.45-0.2 micrometre filter membrane (Millipore, Sydney, Australia or Amicon Sterilet Sydney, Australia) and then stored at 4°C for up to two weeks before use.
- a minimum of six hours or preferably the day before being used the bicarbonate buffered medium is gassed again for two to three minutes with the same gas mixture as above and the protein component is added.
- Tests have been carried out using both mouse embryo development in vitro and with initiation of human pregnancy in an endeavour to discover which components of the T6 medium and the synthetic human tubal fluid according to this invention might be responsible for observed differences in mouse embryo development in vitro and initiation of human pregnancies.
- the results show that for human pregnancy initiation almost three times as many pregnancies occurred when fertilization and culture were carried out using the synthetic human tubal fluid of the present invention over the T6 medium.
- the present invention therefore provides a synthetic human tubal fluid culture medium which is more than just a direct replication of naturally occurring human tubal fluid but has enhanced viability.
- Potassium chloride (Kcl) 4.69 mM
- the invention may be said to reside i n a culture medium for the in vitro fertilization of human oocytes i ncluding sodium potassium i ons wherein the ratio of sodium i ons to potassium ions i s in the range of from 28 to 32.
- the ratio of sodium ions to potassiumi ons is 29.3.
- synthetic human tubal fluid culture medium i s as given in Table 1 below (marked synthetic HTF).
- the medium may be prepared by using rainwater which has been distilled in glass six times.
- the bicarbonate-buffered medium is gassed for a minimum of five minutes with humidified 5% oxygen 5% carbon dioxide 90% nitrogen mixture and sterilized by passage through a 0.45-0.2 micrometre filter membrane (Millipore, Sydney, Australia or Amlcon Sterilet Sydney, Australia) and then stored at 4°C for up to two weeks before use.
- a minimum of six hours or preferably the day before being used the bicarbonate buffered medium is gassed again for two to three minutes with the same gas mixture as above and the protein component is added.
- Tests have been carried out using both mouse embryo development in vitro and with initiation of human pregnancy in an endeavour to discover which components of the T6 medium and the synthetic human tubal fluid according to this invention might be responsible for observed differences in mouse embryo development in vitro and initiation of human pregnancies.
- the results show that for human pregnancy initiation almost three times as many pregnancies occurred when fertilization and culture were carried out
Abstract
A culture medium for in vitro fertilization of human oocytes which comprises: NaCl (96.5 - 106.7 mM) KCl(4.46 - 4.92 mM) MgSO4.7H2O (0.18 - 0.22 mM) KH2PO4 (0.35 - 0.39 mM) CaCl2.2H2O (1.94 - 2.14 mM) NaHCO3 (23.7 - 26.3 mM) Glucose (2.64 - 2.92 mM) Sodium Pyruvate (0.31 - 0.35 mM) Sodium Lactate (20.3 - 22.5 mM) Penicillin (95 - 105 units/ml) Phenol red (5 - 15 micrograms/ml). Culture media wherein the ratio of sodium ions to potassium ions is in the range of from 28 to 32 are preferred.
Description
CULTURE MEDIA FOR IN VITRO FERTILIZATION AND EMBRYO TRANSFER
The invention relates to culture media and more particularly to culture media for in vitro fertilization and embryo transfer.
The process ofi n vitro fertilization of human oocytes, cleavage of embryos and embryo transfer require that a culture medium be used to support the embryo for a period of up to three or four days during the various processes necessary for fertilization and early incubation before embryo transfer and reimplantatlon.
In the natural process of fertilization for a human oocyte the oocyte is supported within the mother within a fluid known as human tubal fluid andi t i s the object of the present invention to provide a culture medium as a synthetic human tubal fluid.
Approximation of the culture conditions as close as possible to those found in the natural environment of the gametes may be most likely to yield the best results. Using this rationale early workers have formulated a culture media similar in biochemical composition to human tubal fluid with varying rates of success. Examples of these include Tyrodes Medium T6, WM1 (Hoppe and Pitts), Modified Earles, and Hams F 10.
One important characteristic of synthetic human tubal fluids appears to the ratio of sodium ions to potassium ions. For natural human tubal fluid this valve is approximately 18. Earlier attempts such as Tyrodes Medium T6 have a value of over 100. We have found that values in between these are most advantageous.
In the present invention we have devised a synthetic culture medium which is believed to approximate human tubal fluid but with desirable additional components and variations in the actual composition, including ratios of concentrations of sodium ions to potassium ions.
In one form therefore the present invention may be said to reside in a culture medium for in vitro fertilization and embryo transfer comprising the following compounds in the following ranges of concentration;
Sodium chloride (NaCI) 96.5 - 106.7 mM Potassium chloride (KCl) 4.46 - 4.92mM
Magnesium sulphate (MgSO47H2O) 0.18 - 0.22mM
Potassium phosphate monobasic (KH2PO4) 0.35 - 0.39mM
Calcium chloride 2 hydrate (CaCl22H2O) 1.94 - 2.14mM
Sodium bicarbonate (NaHCO3) 23.7 - 26.3mM
Glucose 2.64 - 2.92mM
Sodium Pyruvate 0.31 - 0.35mM
Sodium Lactate 20.3 - 22.5mM Penicillin 95 - 105 units/m l
Phenol red 5 - 15 micrograms/m l
In a preferred embodiment of the invention, the ratio of sodium ion concentration to potassium ion concentration is in the range from 28 to 32.
In a further preferred embodiment the ratio of concentrations of sodium ions to potassium ions is 29.3.
In one preferred embodiment of the i nvention the synthetic human tubal fluid may have the following composition;
Sodium Chloride (NaCI) 101.6 mM
Potassium chloride (KCl) 4.69 mM Magnesium sulphate (MgSO47H2O) 0.20mM
Potassium phosphate monobasic (KH2PO4) 0.37mM
Calcium chloride 2 hydrate
(CaCl22H2O) 2.04mM Sodium bicarbonate (NaHCO3) 25.0mM
Glucose 2.78mM
Sodium pyruvate 0.33mM
Sodium lactate 21.4mM
Penicillin 100 units/ml
Phenol red 10 micrograms/ml
In another form the invention may be said to reside in a method of assisting with the in vitro fertilization of human oocytes including the step of handling the human oocytes in a culture medium, the culture medium being comprised of the compounds listed below in the range of compositions listed as follows;
Sodium chloride (NaCI ) 96.5 - 106.7mM
Potassium chloride (KCl ) 4.46 - 4.92mM
Magnesium sulphate (MgSO4 7H2O) 0.18 - 0.22mM
Potassium phosphate monobasic
(KH2PO4) 0.35 - 0.39mM
Calcium chloride 2 hydrate
(CaCl 2 2H2 O) 1.94 - 2.14mM
Sodium bicarbonate (NaHCO3) 23.7 - 26.3mM
Glucose 2.64 - 2.92mM
Sodium pyruvate 0.31 - 0.35mM
Sodium lactate 20.3 - 22.5mM
Penicillin 95 - 105 units/ml
Phenol red 5 - 15 micrograms/ml
In a preferred embodiment of this method of the invention, the ratio of sodium ion concentration to potassium ion concentration is in the range of from 28 to 32.
In a further preferred form of the invention, the ratio concentration of sodium ions to potassium ions is 29.3.
In a further preferred form of the invention, the method includes the step of handling the oocytes in a culture medium comprising;
Sodium Chloride (NaCI) 101.6 mM
Potassium chloride (Kcl) 4.69 mM
Magnesium sulphate (MgSO47H2O) 0.20mM
Potassium phosphate monobasic (KH2PO4) 0.37mM
Calcium chloride 2 hydrate
(CaCl22H2O) 2.04mM
Sodium bicarbonate (NaHCO3) 25.0mM
Glucose 2.78mM
Sodium pyruvate 0.33mM
Sodium lactate 21.4mM
Penicillin 100 units/ml
Phenol red 10 micrograms/ml
In a further form thei nvention may be said to reside in a culture medium for thei n vitro fertilization of human oocytesi ncluding sodium potassiumi ons wherein the ratio of sodium ions to potassium ions is i n the range of from 28 to 32.
In a preferred embodiment of this form of the invention, the ratio of sodium ions to potassium ions is 29.3.
This then generally describes the nature of the present invention and it will be seen that by this invention there is provided a culture medium which is not exactly the same as natural human tubal fluid, but which is capable of supporting in vitro fertilization.
To more clearly assist with the understanding of this invention reference will now be made to a preferred embodiment and tests to determine the efficacy of the preferred embodiment.
In one preferred embodiment synthetic human tubal fluid culture medium is as given in Table 1 below (marked synthetic HTF).
The medium may be prepared by using rainwater which has been distilled in glass six times. The bicarbonate-buffered medium is gassed for a minimum of five minutes with humidified 5% oxygen 5% carbon dioxide
90% nitrogen mixture and sterilized by passage through a 0.45-0.2 micrometre filter membrane (Millipore, Sydney, Australia or Amicon Sterilet Adelaide, Australia) and then stored at 4°C for up to two weeks before use. A minimum of six hours or preferably the day before being used the bicarbonate buffered medium is gassed again for two to three minutes with the same gas mixture as above and the protein component is added.
In a similar way a known culture medium Tyrodes Medium T6 having a composition as given in Table 1 below was also prepared.
T6 FT
l O O l . O O
100 100
Tests have been carried out using both mouse embryo development in vitro and with initiation of human pregnancy in an endeavour to discover which components of the T6 medium and the synthetic human tubal fluid according to this invention might be responsible for observed differences in mouse embryo development in vitro and initiation of human pregnancies. The results show that for human pregnancy initiation almost three times as many pregnancies occurred when fertilization and culture were carried out
using the synthetic human tubal fluid of the present invention over the T6 medium.
In comparison of the two compositions a greatest difference in composition of the two media are their Na+/K+ ratios. We refer to these as sodium/potassium for the rest of the specification.
The ratios of concentrations of sodium ions to potassium ions for these are as follows:
Synthetic Human Tubal Fluid according to the present invention - 29.3
T6 - 105
When media are tested with sodium/potassium leveis varying from 150.5/ 1.42 millimoles to 148.2/5.06 millimoles results showed that there was significant linear and quadratic responses in the number of embryos developing to expanded blastocysts with increasing levels of K+.
In medium containing the potassium levels of T6 medium 75% of the zygotes developed which was significantly fewer than the 95-100% embryos developing when the potassium level was 2.3 to 5.1 millimoles which is the range for the synthetic human tubal fluid of the present invention.
The greatest number of mouse zygotes developing to expanded blastocysts when cultured In synthetic human tubal fluid medium of the present invention compared to T6 medium was paralleled by the three fold increase of the number of pregnancies initiated in those patients whose gametes had been fertilized and cultured in the medium in the present invention rather than T6 medium.
The present invention therefore provides a synthetic human tubal fluid culture medium which is more than just a direct replication of naturally occurring human tubal fluid but has enhanced viability.
Potassium chloride (Kcl) 4.69 mM
Magnesium sulphate (MgSO47H2O) 0.20mM
Potassium phosphate monobasic (KH2PO4) 0.37mM
Calcium chloride 2 hydrate
(CaCl22H2O) 2.04mM
Sodium bicarbonate (NaHCO3) 25.0mM
Glucose 2.78mM
Sodium pyruvate 0.33mM
Sodium lactate 21.4mM
Penicillin 100 units/ml
Phenol red 10 micrograms/ml
In a further form the invention may be said to reside i n a culture medium for the in vitro fertilization of human oocytes i ncluding sodium potassium i ons wherein the ratio of sodium i ons to potassium ions i s in the range of from 28 to 32.
In a preferred embodiment of this form of the invention, the ratio of sodium ions to potassiumi ons is 29.3.
This then generally describes the nature of the present invention and it will be seen that by this invention there is provided a culture medium which i s not exactly the same as natural human tubal fluid, but which is capable of supporting in vitro fertilization.
To more clearly assist with the understanding of this invention reference will now be made to a preferred embodiment and tests to determine the efficacy of the preferred embodiment.
In one preferred embodiment synthetic human tubal fluid culture medium i s as given in Table 1 below (marked synthetic HTF).
The medium may be prepared by using rainwater which has been distilled in glass six times. The bicarbonate-buffered medium is gassed for a minimum of five minutes with humidified 5% oxygen 5% carbon dioxide
90% nitrogen mixture and sterilized by passage through a 0.45-0.2 micrometre filter membrane (Millipore, Sydney, Australia or Amlcon Sterilet Adelaide, Australia) and then stored at 4°C for up to two weeks before use. A minimum of six hours or preferably the day before being used the bicarbonate buffered medium is gassed again for two to three minutes with the same gas mixture as above and the protein component is added.
In a similar way a known culture medium Tyrodes Medium T6 having a composition as given in Table 1 below was also prepared.
T6
HFT
l O O O . O O
100 100
Tests have been carried out using both mouse embryo development in vitro and with initiation of human pregnancy in an endeavour to discover which components of the T6 medium and the synthetic human tubal fluid according to this invention might be responsible for observed differences in mouse embryo development in vitro and initiation of human pregnancies. The results show that for human pregnancy initiation almost three times as many pregnancies occurred when fertilization and culture were carried out
Claims
1. A culture medium for in vitro fertilization of human oocytes comprising:
Sodium chloride (NaCI) 96.5 - 106.7 mM Potassium chloride (KCl) 4.46 - 4.92mM Magnesium sulphate (MgSO47H2O) 0.18 - 0.22mM
Potassium phosphate monobasic (KH2PO4) 0.35 - 0.39mM
Calcium chloride 2 hydrate (CaCl22H2O) 1.94 - 2.14mM
Sodium bicarbonate (NaHCO3) 23.7 - 26.3mM
Glucose 2.64 - 2.92mM
Sodium Pyruvate 0.31 - 0.35mM
Sodium Lactate 20.3 - 22.5mM
Penicillin 95 - 105 units/m l
Phenol red 5 - 15 micrograms/m l
2. A culture medium in Claim 1 wherein the ratio of sodium ion concentration to potassium ion concentration is in the range of from 28 to 32.
3. A culture medium as in Claim 2 wherein the ratio of concentrations of sodium ions to potassium ions is approximately 29.3.
4. A culture medium as in Claim 1 comprising approximately;
Sodium Chloride (NaCI) 101.6 mM Potassium chloride (KCl) 469 mM Magnesium sulphate (MgSO47H2O) 0.20mM
Potassium phosphate monobasic (KH2PO4) 0.37mM
Calcium chloride 2 hydrate
(CaCl22H2O) 2.04mM Sodium bicarbonate (NaHCO3) 25.0mM Glucose 2.78 mM Sodium pyruvate 0.33mM Sodium lactate 21.4mM
Penicillin 100 units/ml
Phenol red 10 micrograms/ml
5. A method of assisting with the in vitro fertilization of human oocytes including the steps of handling the oocytes in a culture medium, the culture medium being of the compounds being listed below in the range of compositions as follows;
Sodium chloride (NaCI) 96.5 - 106.7 mM
Potassium chloride (KCl) 4.46 - 4.92mM
Magnesium sulphate (MgSO47H2O) 0.18 - 0.22mM
Potassium phosphate monobasic (KH2PO4) 0.35 - 0.39mM Calcium chloride 2 hydrate
(CaCl22H2O) 1.94 - 2.14mM
Sodium bicarbonate (NaHCO3) 23.7 - 26.3mM
Glucose 2.64 - 2.92mM
Sodium Pyruvate 0.31 - 0.35mM Sodium Lactate 20.3 - 22.5mM
Penicillin 95 - 105 units/m l
Phenol red 5 - 15 micrograms/m l
6. A method as in Claim 5 wherein the ratio of sodium ion concentration to potassium ion concentration is in the range of from 28 to 32.
7. A method as in Claim 6 wherein the ratio of concentration of sodium ions to potassium ions is approximately 29.3.
8. A method as in Claim 5 wherein the culture medium has a concentration as follows;
Sodium Chloride (NaCI) 101.6 mM
Potassium chloride (Kcl) 4.69 mM Magnesium sulphate (MgSO47H2O) 0.20mM
Potassium phosphate monobasic (KH2PO4) 0.37mM Calcium chloride 2 hydrate
(CaCl22H2O) 2.04mM Sodium bicarbonate (NaHCO3) 25.0mM Glucose 2.78mM
Sodium pyruvate 0.33mM Sodium lactate 21.4mM Penicillin 100 units/ml Phenol red 10 micrograms/ml
9. A culture medium for the in vitro fertilization of human oocytes including sodium and potassium ions wherein the ratio of sodium ions to potassium ions is in the range of from 28 to 32.
10. A culture medium as in Claim 9 wherein the ratio of sodium Ions to potassium ions is approximately 29.3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DK066187A DK66187A (en) | 1985-06-12 | 1987-02-10 | CULTIVATION MEDIA FOR IN VITRO FERTILIZATION AND EMBROIDERY TRANSFER |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AUPH1009 | 1985-06-12 | ||
| AUPH100985 | 1985-06-12 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO1986007377A1 true WO1986007377A1 (en) | 1986-12-18 |
Family
ID=3771143
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/AU1986/000170 WO1986007377A1 (en) | 1985-06-12 | 1986-06-12 | Culture media for in vitro fertilization and embryo transfer |
Country Status (3)
| Country | Link |
|---|---|
| EP (1) | EP0230430A4 (en) |
| DK (1) | DK66187A (en) |
| WO (1) | WO1986007377A1 (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0606847A2 (en) * | 1993-01-07 | 1994-07-20 | National Institute Of Animal Industry, Ministry Of Agriculture, Forestry And Fisheries | Medium for in vitro fertilization and method of in vitro fertilization |
| EP1146837A1 (en) * | 1998-11-30 | 2001-10-24 | IVF Sciences Colorado, Inc. | System and sequential culture media for in vitro fertilization |
| US6586247B1 (en) * | 1999-01-05 | 2003-07-01 | Nipro Corporation | Medium for cultivating animal embryo |
| WO2005063972A1 (en) | 2003-12-30 | 2005-07-14 | Seoul National University Industry Foundation | Embryonic stem cell line and method for preparing the same |
| CN104394689A (en) * | 2011-12-09 | 2015-03-04 | 密苏里大学管理者 | Inorganic pyrophosphate and uses thereof |
| CN111778202A (en) * | 2020-07-10 | 2020-10-16 | 深圳韦拓生物科技有限公司 | Granular cell removing liquid and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4443432A (en) * | 1981-10-05 | 1984-04-17 | Alcon Laboratories, Inc. | Ophthmalic irrigating solution |
| US4473647A (en) * | 1981-02-27 | 1984-09-25 | Amf Inc. | Tissue culture medium |
| WO1986000335A1 (en) * | 1984-06-22 | 1986-01-16 | Veech Richard L | Electrolyte solutions and in vitro use thereof |
-
1986
- 1986-06-12 EP EP19860903165 patent/EP0230430A4/en not_active Withdrawn
- 1986-06-12 WO PCT/AU1986/000170 patent/WO1986007377A1/en not_active Application Discontinuation
-
1987
- 1987-02-10 DK DK066187A patent/DK66187A/en not_active Application Discontinuation
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4473647A (en) * | 1981-02-27 | 1984-09-25 | Amf Inc. | Tissue culture medium |
| US4443432A (en) * | 1981-10-05 | 1984-04-17 | Alcon Laboratories, Inc. | Ophthmalic irrigating solution |
| WO1986000335A1 (en) * | 1984-06-22 | 1986-01-16 | Veech Richard L | Electrolyte solutions and in vitro use thereof |
Non-Patent Citations (5)
| Title |
|---|
| 'Clinical in Vitro Fertilization', Edited by A. TROUNSON and C. WOOD, published 1984 (Springer-Verslag, Berlin) pages 104-105 * |
| Fertility and Sterility, Volume 44, No. 4, issued October 1985, QUINN et al, "Improved Pregnancy Rate in Human in Vitro Fertilization with the Use of a Medium Based on the Composition of Human Tubal Fluid', see pages 493-498 * |
| 'Human Conception in Vitro', Edited by R.G. EDWARDS and J.M. PURDY, published 1982 (Academic Press Inc., London) pages 138-139, 214 * |
| 'In Vitro Fertilization and Embryo Transfer', Edited by A. TROUNSON and C. WOOD, published 1984 (Churchill Livingstone) pages 32-33, 119-120 * |
| See also references of EP0230430A4 * |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0606847A2 (en) * | 1993-01-07 | 1994-07-20 | National Institute Of Animal Industry, Ministry Of Agriculture, Forestry And Fisheries | Medium for in vitro fertilization and method of in vitro fertilization |
| EP0606847A3 (en) * | 1993-01-07 | 1995-04-26 | Nat Inst Animal Ind | Medium for in vitro fertilization and method of in vitro fertilization. |
| EP1146837A1 (en) * | 1998-11-30 | 2001-10-24 | IVF Sciences Colorado, Inc. | System and sequential culture media for in vitro fertilization |
| EP1146837A4 (en) * | 1998-11-30 | 2002-10-31 | Ivf Sciences Colorado Inc | System and sequential culture media for in vitro fertilization |
| US6838235B2 (en) | 1998-11-30 | 2005-01-04 | Vitrolife Group | Methods for in vitro fertilization |
| US6586247B1 (en) * | 1999-01-05 | 2003-07-01 | Nipro Corporation | Medium for cultivating animal embryo |
| AU776033B2 (en) * | 1999-01-05 | 2004-08-26 | Saturo Kaneko | Medium for cultivating animal embryo |
| EP1711599A1 (en) * | 2003-12-30 | 2006-10-18 | Seoul National University Industry Foundation | Embryonic stem cell line and method for preparing the same |
| WO2005063972A1 (en) | 2003-12-30 | 2005-07-14 | Seoul National University Industry Foundation | Embryonic stem cell line and method for preparing the same |
| EP1711599A4 (en) * | 2003-12-30 | 2008-04-16 | Seoul Nat Univ Ind Foundation | Embryonic stem cell line and method for preparing the same |
| US8647872B2 (en) | 2003-12-30 | 2014-02-11 | H. Bion Co., Ltd. | Human embryonic stem cell line prepared by nuclear transfer of a human somatic cell into an enucleated human oocyte |
| US9476064B2 (en) | 2003-12-30 | 2016-10-25 | H Bion Co., Ltd. | Neuro-progenitor produced from an SCNT-derived ES cell |
| CN104394689A (en) * | 2011-12-09 | 2015-03-04 | 密苏里大学管理者 | Inorganic pyrophosphate and uses thereof |
| EP2787810A4 (en) * | 2011-12-09 | 2015-05-27 | Univ Missouri | Inorganic pyrophosphate and uses thereof |
| US10070889B2 (en) | 2011-12-09 | 2018-09-11 | The Curators Of The University Of Missouri | Inorganic pyrophosphate and uses thereof |
| CN111778202A (en) * | 2020-07-10 | 2020-10-16 | 深圳韦拓生物科技有限公司 | Granular cell removing liquid and preparation method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| EP0230430A4 (en) | 1989-03-09 |
| DK66187D0 (en) | 1987-02-10 |
| DK66187A (en) | 1987-03-26 |
| EP0230430A1 (en) | 1987-08-05 |
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