* Corresponding Author
Kemampuan Fertilisasi Spermatozoa Sexing dan Perkembangan Awal Embrio Secara In Vitro pada Sapi
https://doi.org/10.22146/jsv.27562 Alvien Nur Aini(1*), Mohamad Agus Setiadi(2), Ni Wayan Kurniani Karja(3)
(1) Fakultas Kedokteran Hewan, Institut Pertanian Bogor
(2) Fakultas Kedokteran Hewan, Institut Pertanian Bogor
(3) Fakultas Kedokteran Hewan, Institut Pertanian Bogor
(*) Corresponding Author
Abstract
The aim of the present study was to investigate the fertilization ability of bovine oocytes and early bovine embryonic development in vitro, fertilized by frozen X and Y sperm separated by bovine serum albumin (BSA) gradient column. Oocytes were collected from slaughter house ovarian by flushing and slicing technique. Oocytes were than maturated in tissue culture medium (TCM) 199 supplemented with 10 IU/ml pregnant mare’s serum gonadotropin (PMSG), 10 IU/ml human chorionic gonadotropin (hCG) and 10% fetal bovine serum (FBS) for 24 h in 5% CO2 incubator 39oC. Oocytes then fertilized with three kind of different frozen spermatozoa (X,Y and unsexing spermatozoa as control) for 14 h with final concentration 2x106 spermatozoa/mL. Embryos were cultured in
synthetic oviductal fluid (SOF) supplemented with essential and non essential amino acid and 0.3% bovine serum albumin (BSA) for 96 h. Results of the experiments revealed that there was no significant difference (P>0.05) in the
fertilization ability (49.17%; 51.40%; 53.42%) for X, Y and control group, respectively. No significant difference (P>0.05) in the number of embryos development (47.77%; 48.25%; 54.43%) for X, Y and control group, respectively. Furthermore, only small number of embryos could pass development blockade (23.80%; 26.08%; 23.61%) for X, Y and control spermatozoa with statistically no significant difference (P>0.05). It is concluded that sexed spermatozoa separated by BSA gradient column had comparable fertilization ability with unsexing spermatozoa and had ability to supported early embryonic development.
Keywords
Full Text:
PDFReferences
Afiati F. (2004). Proporsi dan karakteristik spermatozoa X dan Y hasil separasi kolom albumin. Media petern 27(1):16-20.
Blondin, P., Beaulieu, M., Fournier, V., Morin, M., Crawford, L., Madan, P and King, W.A.. (2009).
Analysis of bovine sexed sperm for IVF from sorting to the embryo. Theriogenology 71:30-38.
Breitbart H. 2003. Signaling pathways in sperm capacitation and acrosome reaction. Cell Mol Biol . 49(3):1-7.
Dow MP, Bavister BD. 1989. Direct contact is required between serum albumin and hamster spermatozoa for capacitation in vitro. Gamete Res. 23:171-180.
Graf, A., Krebs, S., Heininen-Brown, M., Zakhartchenko,
V., Blum, H. and Wolf, E. (2014a). Genom activation in bovine embryos: review of the literature and new insights from RNA sequencing experiment. Anim Reprod Sci 149:46-58.
Graf, A., Krebs, S., Heininen-Brown, M.,
Zakhartchenko, V., Blum, H. and Wolf, E.
(2014b). Fine mapping of genome activation
in bovine embryos by RNA sequencing. Dev
Biol 111 (11): 4139-4144.
Grant, V.J. and Chamley, L.W. (2007). Sex sorted
sperm and fertility: An alternative view. Biol
reprod 76:184-188.
Jo, H.T., Bang, J.I., Kim, S.S., Choi, B.H., Jin, J.I.,
Kim, H.L., Jung, I.S., Suh, T.K., Ghanem, N.,
Wang, Z. and Kong, I.K. (2014). Production
Alvien Nur Aini et al.
of female bovine embryos with sex-sorted
sperm using intracytoplasmic sperm injection:
Efficiency and in vitro developmental
competence. Theriogenology 81:675-682.
Kaiin, E.M., Said, S. dan Tappa, B. (2008). Kelahiran
anak sapi hasil fertilisasi secara in vitro
dengan sperma hasil pemisahan. Media Petern
(1):22-28.
Lonergan, P. and Fair, T. (2008). In vitro produced
bovine embryos- Dealing with the warts.
Theriogenology 69:17-22.
Lopez, S.R., Souza, J.Cd., Gonzalez, J.Z., Sanchez,
A.D.O., Aguirregomezcorta, J.R., Carvalho,
R.R.de. and Rath, D. (2013). Use of sex sorted
and unsorted frozen/thawed sperm and in vitro
fertilization events in bovine oocytes derived
from ultrasound-guided aspiration. J R Bras
Zootec 42(10):721-727.
Machado, G.M., Carvalho, J.O., Filho, E.S., Caixeta,
E.S., Franco, M.M., Rumpf, R. and Dode,
M.A.N. (2009). Effect of percoll volume,
duration and force of centrifugation, on in vitro
production and sex ratio of bovine embryos.
Theriogenology 71:1289-1297.
Mari, G., Catagnetti, C., Rizzato, G., Mislei, B.,
Lacono, E and Merio, B. (2011). Density
gradient centrifugation of sperm from subfertile
stallion and effect on seminal plasma addition
on fertility. Anim Reprod Sci 126: 96-100.
Meirelles, F.V., Caetano, A.R., Watanabe, Y.F.,
Ripamonte, P., Carambula, S.F., Merighe,
G.K. and Garcia, S.M. (2004). Genome
activation and developmental block in bovine
embryos. Anim Reprod Sci 82-83:13-20.
Morotti, F., Sanches, B.V., Pontes, J.H.F., Basso,
A.C., Siqueira, E.R. and Lisboa, L.A. (2014).
Pregnancy rate and birth rate of calves from a
large-scale IVF program using reverse-sorted
semen in Bos indicus, Bos indicus-taurus and
Bos taurus cattle. Theriogenology 81: 696:701.
Muttaqin, Z., Karja, N.W.K. dan Setiadi, M.A.
(2015). Kemampuan maturasi dan fetilisasi
oosit sapi yang diseleksi menggunakan teknik
pewarnaan Brilliant Cresyl Blue. J Veteriner
(2):242-248.
Nedambale, T.L., Du, F., Yang, X. and Tian, X.C.
(2006). Higher survival rate of vitrified and
thawed in vitro produced blastocysts following
culture in defined medium supplemented with
β-mercaptoethanol. Anim Reprod Sci 93:61-75.
Pellegrino, C.A.G., Morotti. F., Untura, R.M.,
Pontes, J.H.F., Pellegrino, M.F.O., Campolina,
J.P., Seneda, M.M., Barsosa, F.A. and Henry,
M. (2016). Use of sexed sorted semen for
fixed time artificial insemination or fixed tima
embryo transfer of in vitro produced embryos
in cattle. Theriogenology xxx: 1-6.
Setiadi, M.A. dan Karja, N.W.K. (2013). Tingkat
perkembangan awal embrio sapi in vitro
menggunakan media tunggal berbahan dasar
tissue culture medium (TCM) 1999. J Ked
Hewan 7(2):150-154.
Sureka, P., Nilani, K., Eswaramohan, T. and
Balasubramaniam, K. (2013). Sex pre-selection
by quantification of Y chromosome bearing
spermatozoa in goat species. International
Journal of Scientific and Research Publications
(1):1-4.
Suzuki, K., Eriksson B., Shimizu H., Nagai T. and
Rodriguez-Martinez H (2000). Effect of
hyaluronan on monospermic penetration of
porcine oocytes fertilized in vitro. Int J Androl
: 13-21.
Underwood, S.L., Bathgate, R., Maxwell, W.M.C.
and Evans, G. (2010a). Birth of offspring after
artificial insemination of heifers with frozen
thawed, sex sorted, refrozen thawed bull
sperm. Anim Reprod Sci 118:171–175.
Underwood, S.L., Bathgate, R., Pereira, D.C.,
Castro, A., Thomson, P.C., Maxwell, W.M.C.
and Evans, G. (2010b). Embryos production
after in vitro fertilization with frozen thawed,
sex sorted, re frozen thawe bull sperm.
Theriogenology 73:97-102.
Villamil, P.R., Wei, H., Moreira, G., Caccia,
M., Taranco, M.F. and Bo, G.A. (2012).
Fertilization rate and in vitro embryo
production using sexed or non sexed semen
selected with silane-coated silica colloid or
percoll. Theriogenology 78:165-171.
Yan, J., Feng, H.L., Chen, Z.J., Jingmei, H., Xuan, G.
and Yingying, Q. (2006). Influence of swim-up
on the ratio of X and Y bearing spermatozoa.
Eur J Obstet Gynecol Reprod Biol. 129: 150-
DOI: https://doi.org/10.22146/jsv.27562
Article Metrics
Abstract views : 8622
| 
views : 13118
Refbacks
- There are currently no refbacks.
Copyright (c) 2017 Jurnal Sain Veteriner
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
Jurnal Sain Veteriner Indexed by
Copyright of JSV (Jurnal Sain Veteriner) ISSN 0126-0421 (print), ISSN 2407-3733 (online).
Fakultas Kedokteran Hewan, Universitas Gadjah Mada
Jl. Fauna No.2, Karangmalang, Yogyakarta
Phone: 0274-560862
Fax: 0274-560861
Email: jsv_fkh@ugm.ac.id
HP. 0895363078367
Jurnal Sain Veteriner is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.