Bioengineering Approaches to Improve In Vitro Performance of Prepubertal Lamb Oocytes
Juvenile in vitro embryo technology (JIVET) provides exciting opportunities in animal reproduction by reducing the generation intervals. Prepubertal oocytes are also relevant models for studies on oncofertility. However, current JIVET efficiency is still unpredictable, and further improvements are n...
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doaj-510dfa3b29aa45edb1962cf47dbf0f2f2021-06-30T23:51:03ZengMDPI AGCells2073-44092021-06-01101458145810.3390/cells10061458Bioengineering Approaches to Improve In Vitro Performance of Prepubertal Lamb OocytesAntonella Mastrorocco0Ludovica Cacopardo1Daniela Lamanna2Letizia Temerario3Giacomina Brunetti4Augusto Carluccio5Domenico Robbe6Maria Elena Dell’Aquila7Faculty of Veterinary Medicine, University of Teramo, Loc. Piano d’Accio, 64100 Teramo, ItalyResearch Centre E. Piaggio, University of Pisa, Largo Lucio Lazzarino 1, 56122 Pisa, ItalyDepartment of Biosciences, Biotechnologies & Biopharmaceutics, University of Bari Aldo Moro, Via Edoardo Orabona, 70125 Bari, ItalyDepartment of Biosciences, Biotechnologies & Biopharmaceutics, University of Bari Aldo Moro, Via Edoardo Orabona, 70125 Bari, ItalyDepartment of Biosciences, Biotechnologies & Biopharmaceutics, University of Bari Aldo Moro, Via Edoardo Orabona, 70125 Bari, ItalyFaculty of Veterinary Medicine, University of Teramo, Loc. Piano d’Accio, 64100 Teramo, ItalyFaculty of Veterinary Medicine, University of Teramo, Loc. Piano d’Accio, 64100 Teramo, ItalyDepartment of Biosciences, Biotechnologies & Biopharmaceutics, University of Bari Aldo Moro, Via Edoardo Orabona, 70125 Bari, ItalyJuvenile in vitro embryo technology (JIVET) provides exciting opportunities in animal reproduction by reducing the generation intervals. Prepubertal oocytes are also relevant models for studies on oncofertility. However, current JIVET efficiency is still unpredictable, and further improvements are needed in order for it to be used on a large-scale level. This study applied bioengineering approaches to recreate: (1) the three-dimensional (3D) structure of the cumulus–oocyte complex (COC), by constructing—via bioprinting technologies—alginate-based microbeads (COC-microbeads) for 3D in vitro maturation (3D-IVM); (2) dynamic IVM conditions, by culturing the COC in a millifluidic bioreactor; and (3) an artificial follicular wall with basal membrane, by adding granulosa cells (GCs) and type I collagen (CI) during bioprinting. The results show that oocyte nuclear and cytoplasmic maturation, as well as blastocyst quality, were improved after 3D-IVM compared to 2D controls. The dynamic 3D-IVM did not enhance oocyte maturation, but it improved oocyte bioenergetics compared with static 3D-IVM. The computational model showed higher oxygen levels in the bioreactor with respect to the static well. Microbead enrichment with GCs and CI improved oocyte maturation and bioenergetics. In conclusion, this study demonstrated that bioengineering approaches that mimic the physiological follicle structure could be valuable tools to improve IVM and JIVET.https://www.mdpi.com/2073-4409/10/6/1458prepubertal lamb oocyteJIVET3D in vitro maturationmillifluidic bioreactorbioprintinggranulosa cells |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Antonella Mastrorocco Ludovica Cacopardo Daniela Lamanna Letizia Temerario Giacomina Brunetti Augusto Carluccio Domenico Robbe Maria Elena Dell’Aquila |
spellingShingle |
Antonella Mastrorocco Ludovica Cacopardo Daniela Lamanna Letizia Temerario Giacomina Brunetti Augusto Carluccio Domenico Robbe Maria Elena Dell’Aquila Bioengineering Approaches to Improve In Vitro Performance of Prepubertal Lamb Oocytes Cells prepubertal lamb oocyte JIVET 3D in vitro maturation millifluidic bioreactor bioprinting granulosa cells |
author_facet |
Antonella Mastrorocco Ludovica Cacopardo Daniela Lamanna Letizia Temerario Giacomina Brunetti Augusto Carluccio Domenico Robbe Maria Elena Dell’Aquila |
author_sort |
Antonella Mastrorocco |
title |
Bioengineering Approaches to Improve In Vitro Performance of Prepubertal Lamb Oocytes |
title_short |
Bioengineering Approaches to Improve In Vitro Performance of Prepubertal Lamb Oocytes |
title_full |
Bioengineering Approaches to Improve In Vitro Performance of Prepubertal Lamb Oocytes |
title_fullStr |
Bioengineering Approaches to Improve In Vitro Performance of Prepubertal Lamb Oocytes |
title_full_unstemmed |
Bioengineering Approaches to Improve In Vitro Performance of Prepubertal Lamb Oocytes |
title_sort |
bioengineering approaches to improve in vitro performance of prepubertal lamb oocytes |
publisher |
MDPI AG |
series |
Cells |
issn |
2073-4409 |
publishDate |
2021-06-01 |
description |
Juvenile in vitro embryo technology (JIVET) provides exciting opportunities in animal reproduction by reducing the generation intervals. Prepubertal oocytes are also relevant models for studies on oncofertility. However, current JIVET efficiency is still unpredictable, and further improvements are needed in order for it to be used on a large-scale level. This study applied bioengineering approaches to recreate: (1) the three-dimensional (3D) structure of the cumulus–oocyte complex (COC), by constructing—via bioprinting technologies—alginate-based microbeads (COC-microbeads) for 3D in vitro maturation (3D-IVM); (2) dynamic IVM conditions, by culturing the COC in a millifluidic bioreactor; and (3) an artificial follicular wall with basal membrane, by adding granulosa cells (GCs) and type I collagen (CI) during bioprinting. The results show that oocyte nuclear and cytoplasmic maturation, as well as blastocyst quality, were improved after 3D-IVM compared to 2D controls. The dynamic 3D-IVM did not enhance oocyte maturation, but it improved oocyte bioenergetics compared with static 3D-IVM. The computational model showed higher oxygen levels in the bioreactor with respect to the static well. Microbead enrichment with GCs and CI improved oocyte maturation and bioenergetics. In conclusion, this study demonstrated that bioengineering approaches that mimic the physiological follicle structure could be valuable tools to improve IVM and JIVET. |
topic |
prepubertal lamb oocyte JIVET 3D in vitro maturation millifluidic bioreactor bioprinting granulosa cells |
url |
https://www.mdpi.com/2073-4409/10/6/1458 |
work_keys_str_mv |
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