Development of caprine chimeric embryos reconstructed through ES-tetraploid complementation assay
203 / 95
Authors
-
JUHI PATHAK
PhD Scholar, ICAR-Central Institute for Research on Goats, Makhdoom, Mathura, Uttar Pradesh 281 122 India
-
S D KHARCHE
Principal Scientist, ICAR-Central Institute for Research on Goats, Makhdoom, Mathura, Uttar Pradesh 281 122 India
-
ANJANA GOEL
Associate Professor, Department of Biotechnology, Institute of Applied Science and Humanities, GLA
University, Mathura, Uttar Pradesh
Keywords:
Caprine, Chimeric embryos, Culture media, ES-tetraploid complementation assayAbstract
The aim of the present study was to evaluate the development of caprine chimeric embryos in different culture media, viz. RVCL and mCR2aa + 10% FBS. Inner cell mass (ICM) from hatched blastocysts of parthenogenetic activated embryos were used to produce ES cell-like cells (103) while 4-cell embryos obtained from IVF were utilized to produce tetraploid embryos (231). The aggregates prepared were randomly divided into 2 groups, viz. Gr 1 (RVCL) (53) and Gr 2 (mCR2aa +10% FBS) (50) followed by culture in humidified atmosphere of 5% CO2 at 38.5°C in a CO2 incubator. Outcome measures were aggregation, 8–16 cell, morula and blastocyst formation. The percentage of aggregation, 8–16 cell, morula and blastocyst in Gr 1 (RVCL) was 83.33±6.00%, 42.24±7.71%,29.59±7.64% and 11.49±5.36%, respectively while the percentage of aggregation, 8–16 cell, morula and blastocyst in Gr 2 (mCR2aa +10% FBS) was 91.66±4.32%, 54.76±7.75%, 22.32±5.75% and 14.58±5.18%, respectively. In conclusion, both the media supported the development of chimeric embryos up to blastocyst.Downloads
References
Butler J E, Anderson G B, BonDurant R H, Pashen R L and Penedo M C. 1987. Production of ovine chimeras by inner cell mass transplantation. Journal of Animal Science 65: 317–24. DOI: https://doi.org/10.2527/jas1987.651317x
Gutierrez-Adan A, Lonergan P, Rizos D, Ward F A, Boland M P and Pintado B. 2001. Effect of the in vitro culture system on the kinetics of blastocyst development and sex ratio of bovine embryos. Theriogenology 55: 1117–26. DOI: https://doi.org/10.1016/S0093-691X(01)00471-X
Jia W, Yang W, Lei A, Gao Z, Yang C and Hua J. 2008. A caprine chimera produced by injection of embryonic germ cells into a blastocyst. Theriogenology 69(3): 340–48. DOI: https://doi.org/10.1016/j.theriogenology.2007.08.037
Kharche S, Pathak J, Agarwal S, Kushwah B and Sikarwar A K S. 2016. Effect of Ca ionophore on blastocyst production following intracytoplasmic sperm injection in caprine oocytes. Reproduction Domestic Animal 51: 611–17. DOI: https://doi.org/10.1111/rda.12701
Kharche S D, Goel A K, Jindal S K, Goel P and Jha B K. 2011. Birth of twin kids following transfer of in vitro produced caprine embryos. Indian Journal of Animal Sciences 81: 1132– 34.
Nagashima H, Giannakis C, Ashman R J and Nottle M B. 2004. Sex differentiation and germ cell production in chimeric pigs produced by inner cell mass injection into blastocysts. Biology of Reproduction 70(3): 702–07. DOI: https://doi.org/10.1095/biolreprod.103.022681
Pathak J, Kharche S D and Goel A. 2017. Effects of different activation protocols on cleavage rate and blastocyst production of caprine oocytes. Iranian Journal of Veterinary Sciences 18(4): 243–48.
Razza E M, Satrapa R A, Emanuelli I P, Barros C M and Nogueira M F G. 2016. Screening of biotechnical parameters for production of bovine inter-subspecies embryonic chimeras by the aggregation of tetraploid Bos indicus and diploid crossbred Bos taurus embryos. Reproductive Biology 16: 34–40. DOI: https://doi.org/10.1016/j.repbio.2015.11.003
Saito S, Sawai K, Ugai H, Moriyasu S, Minamihashi A and Yamamoto Y. 2003. Generation of cloned calves and transgenic chimeric embryos from bovine embryonic stem-like cells. Biochemistry Biophysics Research Communication 309(1): 104–13. DOI: https://doi.org/10.1016/S0006-291X(03)01536-5
Tarkowski A K. 1961. Mouse chimaeras developed from fused eggs. Nature 190: 857–60. DOI: https://doi.org/10.1038/190857a0
Downloads
Submitted
Published
Issue
Section
License
Copyright (c) 2019 The Indian Journal of Animal Sciences
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
The copyright of the articles published in The Indian Journal of Animal Sciences is vested with the Indian Council of Agricultural Research, which reserves the right to enter into any agreement with any organization in India or abroad, for reprography, photocopying, storage and dissemination of information. The Council has no objection to using the material, provided the information is not being utilized for commercial purposes and wherever the information is being used, proper credit is given to ICAR.
How to Cite
