Embryo vitrification is a fundamental practice in assisted reproduction and fertility preservation. A key step of this process is replacing the internal water with cryoprotectants (CPAs) by transferring embryos from an isotonic to a hypertonic solution of CPAs. However, this applies an abrupt osmotic shock to embryos, resulting in molecular damages that have long been a source of concern. In this study, we introduce a standalone microfluidic system to automate the manual process and minimize the osmotic shock applied to embryos. This device provides the same final CPA concentrations as the manual method but with a gradual increase over time instead of sudden increases. Our system allows the introduction of the dehydrating non-permeating CPA, sucrose, from the onset of CPA-water exchange, which in turn reduced the required time of CPA loading for successful vitrification without compromising its outcomes. We compared the efficacy of our device and the conventional manual procedure by studying vitrified–warmed mouse blastocysts based on their re-expansion and hatching rates and transcription pattern of selected genes involved in endoplasmic reticulum stress, oxidative stress, heat shock, and apoptosis. While both groups of embryos showed comparable re-expansion and hatching rates, on-chip loading reduced the detrimental gene expression of cryopreservation. The device developed here allowed us to automate the CPA loading process and push the boundaries of cryopreservation by minimizing its osmotic stress, shortening the overall process, and reducing its molecular footprint.
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Toward embryo cryopreservation-on-a-chip: A standalone microfluidic platform for gradual loading of cryoprotectants to minimize cryoinjuries
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Research Article|
May 18 2021
Toward embryo cryopreservation-on-a-chip: A standalone microfluidic platform for gradual loading of cryoprotectants to minimize cryoinjuries
Pouria Tirgar
;
Pouria Tirgar
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
2
Department of Bioengineering, McGill University
, Montreal, Quebec H3A0B9, Canada
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Fatemeh Sarmadi;
Fatemeh Sarmadi
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
3
Department of Physiology, McGill University
, Montreal, Quebec H3G1Y6, Canada
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Mojgan Najafi;
Mojgan Najafi
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
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Parinaz Kazemi;
Parinaz Kazemi
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
4
Department of Biology, McGill University
, Montreal, Quebec H4A3J1, Canada
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Sina AzizMohseni;
Sina AzizMohseni
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
5
Electrical Engineering Department, University of British Columbia
, Vancouver, British Columbia V6T1Z1, Canada
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Samaneh Fayazi;
Samaneh Fayazi
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
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Ghazaleh Zandi;
Ghazaleh Zandi
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
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Nikta Ziaie;
Nikta Ziaie
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
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Aida Shoushtari Zadeh Naseri;
Aida Shoushtari Zadeh Naseri
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
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Allen Ehrlicher;
Allen Ehrlicher
2
Department of Bioengineering, McGill University
, Montreal, Quebec H3A0B9, Canada
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Mojtaba Dashtizad
Mojtaba Dashtizad
a)
1
Embryo Biotechnology Laboratory (EmBio Lab), Department of Animal Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB)
, Tehran 1497716316, Iran
a)Author to whom correspondence should be addressed: dashtizad@nigeb.ac.ir. Tel.: +98-21-44787323. Fax: +98-21-44780395
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a)Author to whom correspondence should be addressed: dashtizad@nigeb.ac.ir. Tel.: +98-21-44787323. Fax: +98-21-44780395
Biomicrofluidics 15, 034104 (2021)
Article history
Received:
February 10 2021
Accepted:
April 08 2021
Citation
Pouria Tirgar, Fatemeh Sarmadi, Mojgan Najafi, Parinaz Kazemi, Sina AzizMohseni, Samaneh Fayazi, Ghazaleh Zandi, Nikta Ziaie, Aida Shoushtari Zadeh Naseri, Allen Ehrlicher, Mojtaba Dashtizad; Toward embryo cryopreservation-on-a-chip: A standalone microfluidic platform for gradual loading of cryoprotectants to minimize cryoinjuries. Biomicrofluidics 1 May 2021; 15 (3): 034104. https://doi.org/10.1063/5.0047185
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