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The journey of sperm from its origin to the point of ejaculation begins within the seminiferous tubules of the testis. Here, Sertoli cells produce fluid that propels non-motile sperm through a series of conduits, starting with the straight tubules leading to the rete testis. This interconnected network of tubules acts as the initial pathway for sperm, guiding them into the efferent ductules and then into the epididymis for maturation.
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During fertilization, an egg and sperm cell fuse to create a new diploid structure. In humans, the process occurs once the egg has been released from the ovary, and travels into the fallopian tubes. The process requires several key steps: 1) sperm present in the genital tract must locate the egg; 2) once there, sperm need to release enzymes to help them burrow through the protective zona pellucida of the egg; and 3) the membranes of a single sperm cell and egg must fuse, with the sperm...
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The CARD Method for Mouse Sperm Cryopreservation and In Vitro Fertilization Using Frozen-Thawed Sperm.

Toru Takeo1, Jorge Sztein2, Naomi Nakagata2

  • 1Division of Reproductive Engineering, Center for Animal Resources and Development (CARD), Kumamoto University, Kumamoto, Japan. takeo@kumamoto-u.ac.jp.

Methods in Molecular Biology (Clifton, N.J.)
|October 25, 2018
PubMed
Summary
This summary is machine-generated.

This study introduces an improved sperm cryopreservation protocol for mice, significantly boosting the fertility of frozen-thawed sperm, especially for C57BL/6 strains, enabling reliable in vitro fertilization (IVF). This advancement enhances the management of genetically engineered mice.

Keywords:
CryopreservationIn vitro fertilizationMouseRepositoriesSperm

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Area of Science:

  • Reproductive Biology
  • Cryobiology
  • Genetics

Background:

  • Sperm cryopreservation is vital for managing genetically engineered mice.
  • Reduced fertility of frozen-thawed mouse sperm, particularly C57BL/6 strains, hinders in vitro fertilization (IVF).
  • Existing methods struggle with consistent revitalization of frozen-thawed mouse sperm.

Purpose of the Study:

  • To develop an enhanced sperm cryopreservation and IVF protocol for improved fertility of frozen-thawed mouse sperm.
  • To overcome the low fertility issue associated with frozen-thawed C57BL/6 mouse sperm.
  • To establish a robust method for reliable sperm cryopreservation and IVF in mice.

Main Methods:

  • Modified the cryopreservation protocol by adding L-glutamine to the cryoprotectant.
  • Incorporated methyl-β-cyclodextrin into the sperm preincubation medium.
  • Added reduced glutathione to the fertilization medium for IVF.

Main Results:

  • The new protocol significantly enhanced the fertility of frozen-thawed C57BL/6 mouse sperm.
  • Achieved a stable in vitro fertilization (IVF) rate exceeding 80%.
  • Demonstrated improved outcomes for other mouse strains as well.

Conclusions:

  • The developed protocol offers a robust solution for sperm cryopreservation and IVF.
  • This method enhances the reliability of revitalizing mice from frozen sperm.
  • Improves the archiving and distribution systems for genetically engineered mice.