project - Innovative project
In vitro production of bovine embryos (acronym VIVET)
Veise in vitro embrüote tootmine (akronüüm VIVET)
Objectives
The main objective of the VIVET project is to develop an in vitro embryo production (IVP) system suitable for cattle breeding in Estonia. Additional objectives were the detection of apoptotic mechanisms in embryos that trigger programmed death of bovine ova and embryonic cells in vitro, based on their mRNA and miRNA expression profile; and the development of genome-wide NGS (NGS - next generation sequencing) analysis that detect chromosome mutations in bovine IVP blastocysts.
Objectives
VIVET projekti põhieesmärk on Eesti veisekasvatusele sobiva embrüote kehavälise (in vitro) tootmise süsteemi väljaarendamine. Lisaeesmärkideks oli embrüotes apoptootiliste mehhanismide tuvastamine, mis käivitavad veise munarakkude ja embrüorakkude programmeeritud surma in vitro kasvatamisel, nende mRNA ja miRNA ekspressiooni profiili põhjal ning ülegenoomse NGS analüüsi väljatöötamine, mis tuvastab kromosoomide mutatsioonide esinemissageduse veise in vitro toodetud blastotsüstides.
Activities
Transvaginal follicular puncture (OPU procedure) was performed in 166 dairy cows, of which 13.9% were of Estonian Red and 86.1% Estonian Holstein (EHF) breed. On average, 2.6 oocytes were obtained per procedure. A total of 357 blastocysts were produced in the in vitro system (34.2% of the number of oocytes). The embryo transfers were performed in 133 recipients, 39 of which were heifers and 94 were cows. A total of 21 pregnancies were obtained. The best result were obtained after the transfer of expanded and hatched blastocysts (pregnancy rate 30%). Oocyte donors conceived after 1-2 inseminations, rarely 3 inseminations, which do not differ from cows whose ovaries were not punctured.
Eight OPU procedures were performed in Aberdeen-Angus cattle. In total, 44 oocytes were obtained and 10 blastocysts produced.
As a result of the project, an in vitro embryo production technology package was developed, which can be applied to the cattle sector in Estonia to achieve faster breeding objectives and increase the number of genetically valuable animals in the population.
The second result of the project was the development of an innovative method for analyzing chromosomal mutations in bovine embryos produced in vitro. In the first stage of the experiments, eleven in vitro embryos were sequenced. In a second step, a total of 12 embryos were analyzed, with two samples of each embryo being used (cell sample or biopsy and remaining embryo from biopsy). Analysis of the results showed that with one exception the chromosomal copy number of the embryos and their biopsies was normal. The result shows that the genome-wide screening method we have developed allows us to identify embryos with chromosomal mutations before transplantation, using an embryo biopsy for the analysis. Transplantation of embryos without chromosome mutations will help to increase the likelihood of pregnancy.
Activities
Transvaginaalne folliikulipunktsioon (OPU protseduur) viidi läbi 166 piimalehmal, kellest 13,9% olid eesti punast ja 86,1% eesti holsteini (EHF) tõugu. Keskmiselt saadi ühel protseduuril 2,6 munarakku looma kohta. In vitro süsteemis toodeti kokku 357 blastotsüsti arengujärgus embrüot (34,2% munarakkude arvust). Embrüosiirdamist tehti 133 retsipiendile, kellest 39 olid mullikad ja 94 lehmad. Tiinusi saadi kokku 21. Parima tulemuse andis laienenud või koorunud bastotsüstide siirdamine (tiinestus 30% retsipientidest). Munarakudoonorid ise tiinestusid pärast punktsioonide lõppemist 1-2, harva 3 seemenduse järel, mis ei erine lehmadest, kelle munasarju ei punkteeritud.
Aberdiin-anguse tõugu lihaveistele tehti kaheksa OPU protseduuri. Kokku punkteeriti 117 folliikulit, saadi 44 munarakku ja neist arenes 10 blastotsüsti.
Projekti tulemusena töötati välja veise embrüote in vitro tootmise tehnoloogia pakett, mida on võimalik rakendada kogu veisekasvatussektoris aretuseesmärkide kiiremaks saavutamiseks ja geneetiliselt väärtuslike loomade arvu suurendamiseks populatsioonis.
Projekti teise tulemusena arendati välja innovaatiline meetod, mille abil saab analüüsida kromosoommutatsioone veise in vitro toodetud embrüotes. Katsete esimeses etapis sekveneeriti ülegenoomselt (NGS - next generation sequencing) üksteist in vitro embrüot. Teises etapis analüüsiti kokku 12 embrüot, kusjuures analüüsiks kasutati igast embrüost kahte proovi (rakuproovi ehk bioptaati ja biopsiast alles jäänud embrüot). Tulemuste analüüs näitas, et ühe erandiga oli embrüote ja nende bioptaatide kromosomaalne koopiaarv normaalne. Tulemus näitab, et meie välja arendatud ülegenoomne analüüs võimaldab enne siirdamist kindlaks teha kromosoommutatsioonidega embrüod, kasutades analüüsiks embrüo bioptaati. Ilma kromosoommutatsioonideta embrüote siirdamine aitab tõsta tiinestumise tõenäosust.
Project details
- Main funding source
- Rural development 2014-2020
- Agricultural sectors
- Dairy cattle
€ 392222
Total budget
Total contributions including EU funding.
€ 349726
EU contribution
Any type of EU funding.
Project keyword
Resources
Contacts
Tartu Agro Ltd. Contact person: Aavo Mölder, Estonia, Tähtvere vald, Ilmatsalu, Pargi tee 1, 61401,. tel: +372 7 499272, e-mail: info@tartuagro.ee
Project coordinator
-
Estonian University of Life Sciences
Project coordinator
Project partners
-
The Competence Centre on Health Technologies
Project partner