CN1048168A - 牛x精子分离方法及其分离装置 - Google Patents

牛x精子分离方法及其分离装置 Download PDF

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Publication number
CN1048168A
CN1048168A CN 89105110 CN89105110A CN1048168A CN 1048168 A CN1048168 A CN 1048168A CN 89105110 CN89105110 CN 89105110 CN 89105110 A CN89105110 A CN 89105110A CN 1048168 A CN1048168 A CN 1048168A
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sperm
separator
seminal fluid
electrode
separation method
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苏钟浦
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China Medical University
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China Medical University
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Abstract

本发明提供一种牛X精子分离方法及其分离装 置。该方法可采用鲜精液或冷冻精液进行分离,所用 分离装置是由两端套有电极的电泳池和电极外周缠 有经支持液浸湿的药用纱布卷构成。接通直流电源 1小时后,收集阳极处的精液,即分离出X精子。这 种分离方法分离率高,特别是该方法使用的分离装置 操作简便,所用时间短,分离出的X精子活力好。

Description

当前,为提高奶牛的繁殖率,迅速建立母畜繁殖群,增加牛奶及其奶制品的产量,奶牛繁殖的性别控制已成为畜牧业中主要研究的课题了。由于性别决定的主要因素是性染色体,含X染色体的卵子与含Y或X染色体的精子结合决定子代性别,所以在控制子代性别方面采取了一些相应的措施,如调节饲料、改变母牛生殖道的PH值、掌握人工授精时间等。但是,由于这些措施在目前暂无充分的理论依据,故未能达到预期的效果。为了能在人工授精时控制胎畜的性别,必须使精液中X精子与Y精子分离。本发明者曾针对牛精子性别预测方法和鉴别方法进行了研究(见辽宁畜牧兽医1985年第1期和第3期),提出了简便易行的鉴别方法,并利用电泳法进行了分离牛X及Y精子的初步研究,自行设计的电泳装置虽然取得了较为满意的分离效果,在阳极侧的精液中X精子占90.36%,但因分离出的X精子活力较差,故影响了该装置的推广应用。
本发明的目的是提供一种牛X精子的分离方法及其分离装置,该方法可采用鲜精液或冷冻精液进行分离,分离率高。特别是该方法所使用的分离装置操作简便,所用时间短,分离出的X精子活力好。
本发明的目的是这样实现的:首先将鲜精液装在分离装置的电泳池中,接通直流电源,在电压为3V、电流为40~100μA的条件下,通电1小时后,收集阳极处的精液,即分离出X精子。该方法所用的分离装置的结构是:放置在电泳平台上的电泳池两端各套有一螺旋形电极,电极外周缠有经支持液浸湿的药用纱布卷,两电极分别与直流电源的正、负极相接,电泳池采用可导电的网眼形半透膜袋。
由于本发明的分离方法,采用的电泳池的结构不仅可防止精液与电极直接接触,而且能维持精子的能量来源,不影响精子的运动轨迹,因此,精子的分离率高,分离后的精子活力好。该分离装置结构简单,容易操作,分离时间短,便于携带,特别适于鲜精液的分离,故极易推广应用。
以下结合附图和实施例对本发明作进一步描述。
图1是本发明分离装置一种具体结构的示意图。
图2是图1的A向视图。
根据图1~2详细说明本发明的分离方法及其分离装置的具体结构。该分离装置可封装在一个容器中。其电泳平台6上放置一两端各套有一螺旋形金属钛丝电极2、5的电泳池4。分别与直流电源9正、负极相接的电极2、5的外周缠有经支持液浸湿的药用纱布卷1。该支持液选用可保持渗压,能维持精子能量来源的溶液。本实施例采用浓度为2.9%的柠檬酸钠溶液作为支持液。电泳池4采用可导电的网眼形半透膜袋。操作时,取鲜精液4毫升(取自黑龙江省佳木斯国营农场总局畜牧兽医总站的黑白花种公牛,牛号991),原液活力0.8,密度13亿,X精子所占比例为53.05%。将鲜精液直接置于上述的分离装置中的电泳池内,接通电压为3V的直流电源9和照明灯10的电源,调整电阻箱8使通过的电流为80μA。保持电泳池温度(温度计3显示为37℃),在电流表7的指示为80μA的条件下,通电1小时,断电后收集阳极处的精液得1.1毫升。再将该精液加稀释液3.9毫升,按常规方法制备成冷冻精液颗粒。取出一棵冷冻颗粒,检查活力及X精子所占比例为:活力0.6,X精子占总精子的89.14%,采用本发明的方法分离精液中的X精子,在同样条件下,经十次实验所得结果的平均值可达91.79±6.4,活力为0.68±0.18。如果采用冷冻后的精液分离X精子,应优先选用原料组分体积比为乳糖∶蛋黄∶丙三醇=15∶4∶1的稀释液,稀释比为1∶3即可进行分离。

Claims (2)

1、一种牛X精子分离方法,其特征是将鲜精液或冷冻精液,置于分离装置中,接通直流电源,在电压为3V、电流为40~100μA的条件下,通电1小时后,收集阳极处的精液,即分离出X精子。
2、一种根据权利要求1所述方法用的分离装置,其特征是放置在电泳平台上的电泳池两端各套有一螺旋形电极,电极外周缠有经支持液浸湿的药用纱布卷,两电极分别与直流电源的正、负极相接,电泳池采用可导电的网眼形半透膜袋。
CN 89105110 1989-06-19 1989-06-19 牛x精子分离方法及其分离装置 Pending CN1048168A (zh)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004046712A3 (en) * 2002-11-20 2004-07-22 Univ Virginia Isolation of sperm cells from other biological materials using microfabricated devices and related methods thereof
CN100383237C (zh) * 2004-03-12 2008-04-23 中国科学院昆明动物研究所 大额牛精液低温冷冻保存方法
US8088265B2 (en) * 2003-05-15 2012-01-03 NuSep Holdings Ltd. Cell separation

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004046712A3 (en) * 2002-11-20 2004-07-22 Univ Virginia Isolation of sperm cells from other biological materials using microfabricated devices and related methods thereof
US8088265B2 (en) * 2003-05-15 2012-01-03 NuSep Holdings Ltd. Cell separation
CN100383237C (zh) * 2004-03-12 2008-04-23 中国科学院昆明动物研究所 大额牛精液低温冷冻保存方法

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