Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for Fertilizing Oocytes
Successful in vitro fertilization (IVF) in mice has been achieved using spermatozoa at concentrations specifically optimized for the experimental conditions, such as species and source of spermatozoa. Although IVF in mice is mostly performed using about 80–500 µl drops, it is expected that the numb...
| Main Authors: | , , , , |
|---|---|
| Format: | Online |
| Language: | English |
| Published: |
The Society for Reproduction and Development
2014
|
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4085382/ |
| id |
pubmed-4085382 |
|---|---|
| recordtype |
oai_dc |
| spelling |
pubmed-40853822014-07-08 Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for Fertilizing Oocytes HASEGAWA, Ayumi MOCHIDA, Keiji TOMISHIMA, Toshiko INOUE, Kimiko OGURA, Atsuo Original Article Successful in vitro fertilization (IVF) in mice has been achieved using spermatozoa at concentrations specifically optimized for the experimental conditions, such as species and source of spermatozoa. Although IVF in mice is mostly performed using about 80–500 µl drops, it is expected that the number of spermatozoa used for insemination can be reduced by decreasing the size of the IVF drops. The present study was undertaken to examine the extent to which the number of spermatozoa used for IVF could be reduced by using small droplets (1 µl). We devised the experimental parameters using frozen–thawed spermatozoa from C57BL/6 mice in anticipation of broader applications to other mouse facilities. We found that as few as 5 spermatozoa per droplet could fertilize oocytes (1 or 3 oocytes per droplet), although the fertilization rates were low (13–15%). Practical fertilization rates (> 40%) could be achieved with frozen-thawed C57BL/6J spermatozoa, which are sensitive to cryopreservation, when 20 sperm per droplet were used to inseminate 3 oocytes. Even with spermatozoa from a very poor quality suspension (10% motility), about 25% of oocytes were fertilized. Our calculations indicate that the number of inseminated spermatozoa per oocyte can be reduced to 1/96–1/240 by this method. In two separate embryo transfer experiments, 60% and 47%, respectively, of embryos developed to term. Our microdroplet IVF method may be particularly advantageous when only a limited number of motile spermatozoa are available because of inadequate freezing-thawing or genetic reasons. The Society for Reproduction and Development 2014-02-28 2014-06 /pmc/articles/PMC4085382/ /pubmed/24583808 http://dx.doi.org/10.1262/jrd.2013-136 Text en ©2014 Society for Reproduction and Development http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
HASEGAWA, Ayumi MOCHIDA, Keiji TOMISHIMA, Toshiko INOUE, Kimiko OGURA, Atsuo |
| spellingShingle |
HASEGAWA, Ayumi MOCHIDA, Keiji TOMISHIMA, Toshiko INOUE, Kimiko OGURA, Atsuo Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for Fertilizing Oocytes |
| author_facet |
HASEGAWA, Ayumi MOCHIDA, Keiji TOMISHIMA, Toshiko INOUE, Kimiko OGURA, Atsuo |
| author_sort |
HASEGAWA, Ayumi |
| title |
Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for
Fertilizing Oocytes |
| title_short |
Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for
Fertilizing Oocytes |
| title_full |
Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for
Fertilizing Oocytes |
| title_fullStr |
Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for
Fertilizing Oocytes |
| title_full_unstemmed |
Microdroplet In Vitro Fertilization Can Reduce the Number of Spermatozoa Necessary for
Fertilizing Oocytes |
| title_sort |
microdroplet in vitro fertilization can reduce the number of spermatozoa necessary for
fertilizing oocytes |
| description |
Successful in vitro fertilization (IVF) in mice has been achieved using spermatozoa at concentrations
specifically optimized for the experimental conditions, such as species and source of spermatozoa. Although IVF in mice is
mostly performed using about 80–500 µl drops, it is expected that the number of spermatozoa used for insemination can be
reduced by decreasing the size of the IVF drops. The present study was undertaken to examine the extent to which the number
of spermatozoa used for IVF could be reduced by using small droplets (1 µl). We devised the experimental parameters using
frozen–thawed spermatozoa from C57BL/6 mice in anticipation of broader applications to other mouse facilities. We found that
as few as 5 spermatozoa per droplet could fertilize oocytes (1 or 3 oocytes per droplet), although the fertilization rates
were low (13–15%). Practical fertilization rates (> 40%) could be achieved with frozen-thawed C57BL/6J spermatozoa, which
are sensitive to cryopreservation, when 20 sperm per droplet were used to inseminate 3 oocytes. Even with spermatozoa from a
very poor quality suspension (10% motility), about 25% of oocytes were fertilized. Our calculations indicate that the number
of inseminated spermatozoa per oocyte can be reduced to 1/96–1/240 by this method. In two separate embryo transfer
experiments, 60% and 47%, respectively, of embryos developed to term. Our microdroplet IVF method may be particularly
advantageous when only a limited number of motile spermatozoa are available because of inadequate freezing-thawing or genetic
reasons. |
| publisher |
The Society for Reproduction and Development |
| publishDate |
2014 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4085382/ |
| _version_ |
1613109458418794496 |