Semen parameters and fertility potency of a cloned water buffalo (Bubalus bubalis) bull produced from a semen-derived epithelial cell

Monika Saini; Suman Sheoran; Kennady Vijayalakshmy; Rasika Rajendran; Dharmendra Kumar; Pradeep Kumar; Rakesh Kumar Sharma; Rajesh Kumar; Tushar K. Mohnaty; Naresh L. Selokar; Prem Singh Yadav

doi:10.1371/journal.pone.0237766

Semen parameters and fertility potency of a cloned water buffalo (Bubalus bubalis) bull produced from a semen-derived epithelial cell

Monika Saini, Suman Sheoran, Kennady Vijayalakshmy, Rasika Rajendran, Dharmendra Kumar, Pradeep Kumar, Rakesh Kumar Sharma, Rajesh Kumar, Tushar K. Mohnaty, Naresh L. Selokar, Prem Singh Yadav

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Abstract

Semen contains epithelial cells that can be cultured in vitro. For somatic cell nuclear transfer applications, it is essential to know whether clone(s) produced from semen-derived epithelial cells (SedECs) are healthy and reproductively competent. In this study, the semen and fertility profile of a cloned bull (C1) that was produced from a SedEC were compared with its donor (D1) and with two cloned bulls (C2, C3) that were produced from commonly used skin-derived fibroblast cells (SkdFCs). We observed variations in some fresh semen parameters (ejaculated volume and mass motility), frozen-thawed sperm parameters (plasma membrane integrity, and computer-assisted semen analysis (CASA) indices), but values are within the normal expected range. There was no difference in sperm concentration of ejaculated semen and frozen-thawed semen parameters which include sperm motility, percentage of live and normal morphology sperm, and distance traveled through oestrus mucus. Following in vitro fertilization (IVF) experiments, zygotes from C1 had higher (P < 0.05) cleavage rates (81%) than C2, C3, and D1 (71%, 67%, and 75%, respectively); however, blastocyst development per cleaved embryo and quality of produced blastocysts did not differ. The conception rate of C1 was 46% (7/15) and C2 was 50% (8/15) following artificial insemination with frozen-thawed semen. Established pregnancies resulted in births of 7 and 6 progenies sired by C1 and C2, respectively, and all calves show no signs of phenotypical abnormalities. These results showed that semen from a cloned bull derived from SedECs is equivalent to semen from its donor bull and bulls cloned from SkdFCs.

Original language	English
Article number	e0237766
Journal	PLoS ONE
Volume	15
Issue number	8 August 2020
DOIs	https://doi.org/10.1371/journal.pone.0237766
State	Published - Aug 2020
Externally published	Yes

Bibliographical note

Publisher Copyright:
© 2020 Saini et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding

This study was funded by Indian Council of Agricultural Research, New Delhi (NASF/GTR-7004/2018-19/184) to PSY and Science and Engineering Research Board, New Delhi (YSS/ 2014/000278) to NLS. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We are thankful to Director ICAR-Central Institute for Research on Buffaloes for providing necessary facilities to conduct this study. We thank Prof. Charles R. Long, Texas A&M University, USA, for his valuable comments on the manuscript draft.

Funders	Funder number
Science and Engineering Research Board, New Delhi	YSS/ 2014/000278
Texas A and M University
Indian Council of Agricultural Research	NASF/GTR-7004/2018-19/184

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Saini, M., Sheoran, S., Vijayalakshmy, K., Rajendran, R., Kumar, D., Kumar, P., Sharma, R. K., Kumar, R., Mohnaty, T. K., Selokar, N. L., & Yadav, P. S. (2020). Semen parameters and fertility potency of a cloned water buffalo (Bubalus bubalis) bull produced from a semen-derived epithelial cell. PLoS ONE, 15(8 August 2020), Article e0237766. https://doi.org/10.1371/journal.pone.0237766

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abstract = "Semen contains epithelial cells that can be cultured in vitro. For somatic cell nuclear transfer applications, it is essential to know whether clone(s) produced from semen-derived epithelial cells (SedECs) are healthy and reproductively competent. In this study, the semen and fertility profile of a cloned bull (C1) that was produced from a SedEC were compared with its donor (D1) and with two cloned bulls (C2, C3) that were produced from commonly used skin-derived fibroblast cells (SkdFCs). We observed variations in some fresh semen parameters (ejaculated volume and mass motility), frozen-thawed sperm parameters (plasma membrane integrity, and computer-assisted semen analysis (CASA) indices), but values are within the normal expected range. There was no difference in sperm concentration of ejaculated semen and frozen-thawed semen parameters which include sperm motility, percentage of live and normal morphology sperm, and distance traveled through oestrus mucus. Following in vitro fertilization (IVF) experiments, zygotes from C1 had higher (P < 0.05) cleavage rates (81%) than C2, C3, and D1 (71%, 67%, and 75%, respectively); however, blastocyst development per cleaved embryo and quality of produced blastocysts did not differ. The conception rate of C1 was 46% (7/15) and C2 was 50% (8/15) following artificial insemination with frozen-thawed semen. Established pregnancies resulted in births of 7 and 6 progenies sired by C1 and C2, respectively, and all calves show no signs of phenotypical abnormalities. These results showed that semen from a cloned bull derived from SedECs is equivalent to semen from its donor bull and bulls cloned from SkdFCs.",

author = "Monika Saini and Suman Sheoran and Kennady Vijayalakshmy and Rasika Rajendran and Dharmendra Kumar and Pradeep Kumar and Sharma, {Rakesh Kumar} and Rajesh Kumar and Mohnaty, {Tushar K.} and Selokar, {Naresh L.} and Yadav, {Prem Singh}",

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AU - Rajendran, Rasika

AU - Kumar, Dharmendra

AU - Kumar, Pradeep

AU - Sharma, Rakesh Kumar

AU - Kumar, Rajesh

AU - Mohnaty, Tushar K.

AU - Selokar, Naresh L.

AU - Yadav, Prem Singh

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N2 - Semen contains epithelial cells that can be cultured in vitro. For somatic cell nuclear transfer applications, it is essential to know whether clone(s) produced from semen-derived epithelial cells (SedECs) are healthy and reproductively competent. In this study, the semen and fertility profile of a cloned bull (C1) that was produced from a SedEC were compared with its donor (D1) and with two cloned bulls (C2, C3) that were produced from commonly used skin-derived fibroblast cells (SkdFCs). We observed variations in some fresh semen parameters (ejaculated volume and mass motility), frozen-thawed sperm parameters (plasma membrane integrity, and computer-assisted semen analysis (CASA) indices), but values are within the normal expected range. There was no difference in sperm concentration of ejaculated semen and frozen-thawed semen parameters which include sperm motility, percentage of live and normal morphology sperm, and distance traveled through oestrus mucus. Following in vitro fertilization (IVF) experiments, zygotes from C1 had higher (P < 0.05) cleavage rates (81%) than C2, C3, and D1 (71%, 67%, and 75%, respectively); however, blastocyst development per cleaved embryo and quality of produced blastocysts did not differ. The conception rate of C1 was 46% (7/15) and C2 was 50% (8/15) following artificial insemination with frozen-thawed semen. Established pregnancies resulted in births of 7 and 6 progenies sired by C1 and C2, respectively, and all calves show no signs of phenotypical abnormalities. These results showed that semen from a cloned bull derived from SedECs is equivalent to semen from its donor bull and bulls cloned from SkdFCs.

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Semen parameters and fertility potency of a cloned water buffalo (Bubalus bubalis) bull produced from a semen-derived epithelial cell

Abstract

Bibliographical note

Funding

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