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


bovine; embryo; in vitro; sexing; spermatozoa

Full Text:

PDF


References

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 : 8600 | views : 13076

Refbacks

  • There are currently no refbacks.




Copyright (c) 2017 Jurnal Sain Veteriner

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

Jurnal Sain Veteriner Indexed by

    CrossrefROADCOREProduct DetailsDESKRIPSI GAMBAR


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.

 

free
web stats View My Stats