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A kind of gamete buffer solution and buffer system thereof

A buffer system and buffer technology, applied in the field of assisted reproduction, can solve the problem of neglecting temperature, and achieve the effect of narrow pH value range, beneficial to embryo formation and development, and low toxicity.

Active Publication Date: 2021-04-06
成都艾伟孚生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although MOPS and HEPES are widely used in the field of assisted reproduction, the influence of temperature on the pKa value of the two and the influence of light on MOPS are often ignored

Method used

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  • A kind of gamete buffer solution and buffer system thereof
  • A kind of gamete buffer solution and buffer system thereof
  • A kind of gamete buffer solution and buffer system thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1 temperature is buffered to the influence of pH

[0044] 1. Effect of temperature on the pKa value of MOPS and HEPES

[0045] Table 1 pKa values ​​of MOPS and HEPES buffer pairs

[0046]

[0047]

[0048] It can be seen from the above table that the influence of temperature on the pKa value of each buffer pair is obvious.

[0049] 2. Changes in pH of medium containing MOPS, HEPES, NaHCO3, MOPS+HEPES mixed buffer

[0050] Take out 500uL of medium containing various buffer solutions from the laboratory incubator and place them at room temperature, read the temperature and pH value every minute within 10 minutes, see the results figure 1 , figure 2 and image 3 and Figure 4 .

[0051]As can be seen from the figure above, the amphoteric buffer MOPS and HEPES can provide a more stable pH value in actual operation, but the buffering capacity of each buffer pair is different. The buffer system mixed with MOPS and HEPES is more stable than MOPS or HEP...

Embodiment 2

[0052] The buffer capacity comparison of embodiment 2 buffer pairs

[0053] The buffering capacity of different concentrations of MOPS, HEPES and MOPS+HEPES buffer was compared by titration. The experiment is as follows:

[0054] 0.1mol / L NaOH and 0.1mol / L HCl were used to titrate 25mL of medium containing various buffers. Experimental results such as Figure 5 , 6 . in Figure 5 The buffering capacity of 10mmol / L HEPES+10mmol / L MOPS buffer mixed with 1:1 was compared with that of 10mmol MOPS and 10mmol HEPES buffer; Figure 6 The buffer capacity of 10mmol / LHEPES+10mmol / LMOPS buffer mixed with 1:1 was compared with that of 20mmol MOPS and 20mmol HEPES buffer.

[0055] Figure 5 It shows that the buffering capacity of the two amphoteric buffers is enhanced after mixing, Figure 6 It shows that the buffer capacity of 10mmol / LHEPES+10mmol / L MOPS mixed at 1:1 is equivalent to the buffer capacity of 20mmol MOPS and 20mmol HEPES buffer, and the combined amphoteric buffer can...

Embodiment 3

[0056] Embodiment 3 Preparation of Gamete Buffer 1

[0057] With ultrapure water as solvent, the formula is as follows:

[0058] 90.24mmol / L sodium chloride, 0.43mmol / L magnesium chloride, 5.013mmol / L potassium chloride, 0.2567mmol / L magnesium sulfate (heptahydrate), 0.30mmol / L calcium pantothenate, 0.3513mmol / L potassium phosphate, 21.0mmol / L calcium lactate, 10mmol / L HEPES (free acid), 10mmol / L MOPS, 4.30mmol / L sodium bicarbonate, 0.3129mmol / L sodium pyruvate, 3mmol / L glucose, 1.06mmol / L alanyl glutamine, 0.10 mmol / L L-taurine, 5ug / mL gentamicin sulfate, 5mg / mL human serum albumin and 0.5mmol / L human non-essential amino acids (glycine, L-alanine, L-proline, L - Serine, L-Asparagine.H2O, L-Aspartic Acid, L-Glutamic Acid).

[0059] Adjust pH7.25-7.35, adjust osmotic pressure 285-295mOsm / kg.

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Abstract

The invention belongs to the field of assisted reproduction, and in particular relates to a gamete buffer. The gamete buffer of the present invention adopts the mixed buffer system of MOPS and HEPES, compared with the common gamete buffer that only uses MOPS or HEPES buffer system, it can control the pH in a narrower range, and reduce the MOPS or HEPES components. concentration can reduce cytotoxicity. Therefore, the gamete buffer solution of the present invention can provide a more stable pH value during gamete collection and preparation, reduce environmental fluctuations, and thus maintain gamete viability. Using the gametes washed with the gamete buffer solution, the blastocyst formation rate is greatly improved in late embryo experiments, and the shape is uniform.

Description

technical field [0001] The invention belongs to the field of assisted reproduction, and in particular relates to a buffer system of a gamete buffer and a gamete buffer prepared therefrom. Background technique [0002] Gamete refers to the mature sex cells produced by the reproductive system when organisms undergo sexual reproduction, referred to as germ cells. Gametes are divided into male gametes and female gametes. The female gametes of animals and plants are usually called eggs (ova, or eggs), while the male gametes are called sperm. [0003] Gamete buffer is a buffer containing antibiotics, basic ions and various energy-rich substances, which is used for washing egg cells and sperm outside the incubator to prepare for IVF (in vitro fertilization). [0004] Since the birth of the field of IVF, scientists have struggled to study healthy embryo implantation, a central goal of reproductive science research. Gamete collection and preparation can be the most stressful step i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0608
Inventor 余裕炉衡钱利刘洪君严飞戴甄
Owner 成都艾伟孚生物科技有限公司
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