119:
during flow cytometry, X spermatozoa fluoresce brighter than Y- spermatozoa. As the spermatozoa pass through the flow cytometer in single file, each spermatozoon is encased by a single droplet of fluid and assigned an electric charge corresponding to its chromosome status (e.g. X-positive charge, Y-negative charge). The stream of X- and Y- droplets is then separated by means of electrostatic deflection and collected into separate collection tubes for subsequent processing.
339:'gates' which the operator places around the total population visible to the machine. In general, the larger the DNA difference between the X and Y chromosome of a species, the easier it is to produce a highly pure population. In sheep and cattle, purities for each sex will usually remain above 90% depending on 'gating', while for humans these may be reduced to 90% for "female" spermatozoa and 70% for "male" spermatozoa.
182:(HA) binding sites on the sperm plasma membrane are an indicator of sperm maturity (Huszar et al., 2003, Yudin et al.,1999). There are two methods based on this fact: physiological intracytoplasmic sperm injection (PICSI), and a sperm slow procedure; both methods require sperm preparation via sperm washing or centrifugation.
338:
While highly accurate, sperm sorting by flow cytometry will not produce two completely separate populations. That is to say, there will always be some "male" sperm among the "female" sperm and vice versa. The exact percentage purity of each population is dependent on the species being sorted and the
95:
is another method used to sort sperm and adaptations of this technique opens new opportunities in sperm sorting. However, because flow cytometry-based sperm sorting often uses fluorescent dyes that often stain DNA, the safety of this technique in human reproductive medicine is a matter of scientific
118:
which binds to the DNA of each spermatozoon. As the X chromosome is larger (i.e. has more DNA) than the Y chromosome, the "female" (X-chromosome bearing) spermatozoa will absorb a greater amount of dye than its male (Y-chromosome bearing) counterpart. As a consequence, when exposed to UV light
321:
There is no country in the world which explicitly permits sex selection for non-medical purposes. There were 31 countries in 2009 which allowed sex selection in case of sex-linked disease risk or other medical purpose. In the US, for humans, the application of sperm sorting in sex selection is
226:. Egg cells are often fertilized by sperm which would have low chance of fertilizing it in natural conditions. Sperm sorting could thus be used to decrease risks associated with assisted reproduction. Additionally, there is ongoing debate about using sperm sorting for choosing the child's sex.
83:
techniques apply a centrifugation step and then sperm is allowed to swim up into a medium, thus enriching a subpopulation of motile sperm. However, use of sperm centrifugation is detrimental to the sperm viability and elicits production of reactive oxygen species. Conventional techniques are
280:(FAO) as a promising way of increasing efficiency of agriculture needed to produce enough food for the growing human population. Utilizing artificial insemination with sorted sperm is seen as a way to create an optimal ratio of male and female calves to increase dairy
334:
or having at least one child (for family balancing). There are currently MicroSort laboratories and collaborating physicians in several countries (some for general purposes, some only offering service in case of genetic disease risks associated with one sex).
242:. It has been verified that sperm sorted using these techniques is of superior quality than unsorted. However, important characteristics of sperm such as DNA integrity remain untested by these conventional methods. New flow-cytometry based techniques such as
918:
De Geyter, C.; Bals-Pratsch, M.; Doeren, M.; Yeung, C. H.; Grunert, J. H.; Bordt, J.; Schneider, H. P.; Nieschlag, E. (1988-11-01). "Human and bovine cervical mucus penetration as a test of sperm function for in-vitro fertilization".
113:
was developed by USDA in conjunction with
Lawrence Livermore National Laboratories, relying on the DNA difference between the X- and Y- chromosomes. Prior to flow cytometric sorting, semen is labeled with a fluorescent dye called
54:(male) chromosome bearing populations based on their difference in DNA content. The resultant 'sex-sorted' spermatozoa are then able to be used in conjunction with other assisted reproductive technologies such as
380:
Rappa, Kari L.; Rodriguez, Harold F.; Hakkarainen, Gloria C.; Anchan, Raymond M.; Mutter, George L.; Asghar, Waseem (2016-09-01). "Sperm processing for advanced reproductive technologies: Where are we today?".
271:
Sperm sorting by flow cytometry is an established technique in veterinary practice, and in the dairy industry most female cows are artificially inseminated with sorted semen to increase the number of female
99:
However, flow cytometry is the only currently used technique able to determine the sex of future progeny by measuring DNA content of individual sperm cells. It evaluates if they contain the larger
276:(using sperm sorting is less common in other species of farm animals, however artificial insemination is common). Artificial insemination of farm animals with sorted sperm is recognized by the
176:
and MACS (Magnetic
Activating Cell Sorting), which in combination with density gradient centrifugation in single sperm preparation protocols results in spermatozoa with superior quality.
109:
79:(in a continuous or discontinuous gradient) can concentrate semen samples with low concentration of sperm, using the density of sperm as a measure of their quality. Similarly, so-called
588:
Johnson, L. A.; Welch, G. R.; Rens, W. (1999-01-01). "The
Beltsville sperm sexing technology: high-speed sperm sorting gives improved sperm output for in vitro fertilization and AI".
318:. It could also have implications on gender equality if parents consistently choose to have a boy as their first-born (first-borns were shown to be more likely to succeed in life).
820:
Said, Tamer; Agarwal, Ashok; Grunewald, Sonja; Rasch, Manja; Baumann, Thomas; Kriegel, Christian; Li, Liang; Glander, Hans-Juergen; Thomas, Anthony J.; Paasch, Uwe (2006-03-01).
330:. The procedure was made available to a limited number of participants each month, in addition to fulfilling certain criteria, such as having a disease with
157:. It is not specific enough to detect individual traits, however. The sperm cells having least DNA damage may subsequently be injected into the egg cell by
724:"Prospective controlled trial of an electrophoretic method of sperm preparation for assisted reproduction: comparison with density gradient centrifugation"
211:
39:. Newly applied methods such as flow cytometry expand the possibilities of sperm sorting and new techniques of sperm sorting are being developed.
203:
310:
in families with a history of these diseases. On the other hand, sperm sorting in humans raises the ethical concerns implicit to the idea of
173:
138:, coupled with magnetic beads. Following the binding of these antibodies, spermatozoa which undergo apoptosis are sorted by applying
771:
Kheirollahi-Kouhestani, M.; Razavi, S.; Tavalaee, M.; Deemeh, M.R.; Mardani, M.; Moshtaghian, J.; Nasr-Esfahani, M.H. (2009-10-01).
243:
126:(MACS) which is routinely applied in assisted reproduction hospitals to sort out sperm with fragmented DNA. This is achieved using
427:
Ribeiro, Sofia C.; Sartorius, Gideon; Pletscher, Flurina; Geyter, Maria de; Zhang, Hong; Geyter, Christian de (September 2013).
650:"Sperm selection using magnetic activated cell sorting (MACS) in assisted reproduction: a systematic review and meta-analysis"
965:
Rath, D.; Barcikowski, S.; de Graaf, S.; Garrels, W.; Grossfeld, R.; Klein, S.; Knabe, W.; Knorr, C.; Kues, W. (2013-01-01).
471:
Paasch, Uwe; Grunewald, Sonja; Glander, Hans-Juergen (2007-01-01). "Sperm selection in assisted reproductive techniques".
822:"Selection of Nonapoptotic Spermatozoa As a New Tool for Enhancing Assisted Reproduction Outcomes: An In Vitro Model1"
42:
It can be used to sort out sperm that are most healthy, as well as for determination of more specific traits, such as
353:
323:
277:
871:"Significance of Reactive Oxygen Species and Antioxidants in Defining the Efficacy of Sperm Preparation Techniques"
223:
158:
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when millions of sperm enter the vagina but only few reach the egg cell and then only one is usually allowed to
648:
Gil, Monica; Sar-Shalom, Valerie; Sivira, Yolisid
Melendez; Carreras, Ramon; Checa, Miguel Angel (2013-03-07).
168:
index the population of sperm could be enriched with spermatozoa with non-fragmented DNA, with techniques like
635:
Seidel GE, Jr., Garner DL. Current status of sexing mammalian spermatozoa. Reproduction 2002;124: 733-743.
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626:
Garner DL, Seidel GE. History of commercializing sexed semen for cattle. Theriogenology 2008;69: 886-895.
303:
234:
Conventional methods of sperm sorting have been widely used to assess quality of sperm before subsequent
1056:
722:
Fleming, S.D.; Ilad, R.S.; Griffin, A-M.G.; Wu, Y.; Ong, K.J.; Smith, H.C.; Aitken, R.J. (2008-09-01).
429:"Isolation of spermatozoa with low levels of fragmented DNA with the use of flow cytometry and sorting"
62:(IVF) to produce offspring of the desired sex - in farming animals but also in human medical practice.
1145:
258:
can significantly improve pregnancy rates in couples with previous assisted reproduction failure.
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235:
207:
55:
773:"Selection of sperm based on combined density gradient and Zeta method may improve ICSI outcome"
326:. After the establishment of the MicroSort technique, it was offered to parents as a part of a
299:
239:
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59:
293:
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709:
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8:
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1016:"Ethical arguments for and against sperm sorting for non-medical sex selection: a review"
219:
161:(ICSI). Many other methods for sperm sorting have been proposed or are currently tested.
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to the sperm suspension. MACS obviates the need for fluorescent DNA binding molecules.
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107:(leading to male progeny). It then allows separation of X and Y sperm. The so-called
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967:"Sex selection of sperm in farm animals: status report and developmental prospects"
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Several methods have been used to sort sperm before the advent of flow cytometry.
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Ribeiro, Sofia; Sartorius, Gideon; Geyter, Christian de (September 2013).
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Kalfoglou, A.L.; Kammersell, M.; Philpott, S.; Dahl, E. (March 2013).
508:"Sperm sorting for selection of healthy sperm: is it safe and useful?"
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Ranking sperm cells could improve the odds of in vitro fertilization.
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By
Courtney Humphries. TechnologyReview. Wednesday, January 21, 2009
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314:. If applied large-scale, it has a potential to elicit a
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and dead spermatozoa from the viable ones. For example,
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Genetics & IVF Institute: MicroSort
Clinical Trial
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470:
122:
Another cytometric technique used in sperm sorting is
721:
214:. This selection is largely circumvented in case of
84:
routinely used in assisted reproductive technology.
1057:"Countries with laws or policies on sex selection"
198:it. The sperm is selected not only by its highest
869:Aitken, R. John; Clarkson, Jane S. (1988-11-12).
1117:
587:
473:Society of Reproduction and Fertility Supplement
298:Choosing the sex of children might help prevent
103:(giving rise to a female offspring) or smaller
868:
654:Journal of Assisted Reproduction and Genetics
130:to surface markers of programmed cell death (
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549:"Sorting of spermatozoa with flow cytometry"
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153:in sperm cells may be detected by using
46:in which spermatozoa are separated into
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31:. Several conventional techniques of
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933:10.1093/oxfordjournals.humrep.a136824
294:Sex selection § Ethical concerns
229:
1096:"About Us | MicroSort International"
921:Human Reproduction (Oxford, England)
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19:is a means of choosing what type of
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218:which leads to higher incidence of
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888:10.1002/j.1939-4640.1988.tb01067.x
506:Caroppo, Ettore (September 2013).
202:but also by other factors such as
110:Beltsfield Sperm Sexing Technology
14:
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638:
461:
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354:Preimplantation genetic diagnosis
300:sex-associated heritable diseases
278:Food and Agriculture Organization
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566:10.1016/j.fertnstert.2013.06.039
525:10.1016/j.fertnstert.2013.06.006
446:10.1016/j.fertnstert.2013.05.030
395:10.1016/j.biotechadv.2016.01.007
224:assisted reproductive techniques
159:intracytoplasmic sperm injection
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1020:Reproductive BioMedicine Online
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164:To select spermatozoa with low
124:magnetic-activated cell sorting
77:Density gradient centrifugation
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839:10.1095/biolreprod.105.046607
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190:Sperm undergoes a process of
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7:
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1033:10.1016/j.rbmo.2012.11.007
598:10.2527/1999.77suppl_2213x
304:Duchene muscular dystrophy
291:
246:staining can discriminate
65:
1055:Darnovsky, Marcy (2009).
666:10.1007/s10815-013-9962-8
590:Journal of Animal Science
322:tightly regulated by the
1062:. geneticsandsociety.org
826:Biology of Reproduction
592:. 77 Suppl 2: 213–220.
553:Fertility and Sterility
512:Fertility and Sterility
433:Fertility and Sterility
236:artificial insemination
208:reactive oxygen species
71:Conventional techniques
56:artificial insemination
383:Biotechnology Advances
240:in-vitro fertilization
216:in-vitro fertilization
60:in-vitro fertilization
790:10.1093/humrep/dep088
741:10.1093/humrep/den330
292:Further information:
254:staining followed by
875:Journal of Andrology
984:10.1530/REP-12-0151
316:sex-ratio imbalance
1131:Fertility medicine
777:Human Reproduction
728:Human Reproduction
230:For general health
155:Raman spectroscopy
1100:www.microsort.com
783:(10): 2409–2416.
734:(12): 2646–2651.
439:(3): 686–694.e4.
262:For sex selection
192:natural selection
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146:Other techniques
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1026:(3): 231–239.
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977:(1): R15–30.
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50:(female) and
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17:Sperm sorting
1151:Reproduction
1103:. Retrieved
1099:
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1064:. Retrieved
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186:Applications
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105:Y chromosome
101:X chromosome
98:
96:discussion.
91:
74:
41:
16:
15:
1094:ThemeFuse.
479:: 515–525.
332:sex linkage
308:haemophilia
1141:Obstetrics
1120:Categories
1105:2016-10-06
1066:2016-10-06
559:(3): e15.
360:References
267:In farming
166:DNA damage
151:DNA damage
134:) such as
128:antibodies
21:sperm cell
993:1741-7899
941:0268-1161
897:1939-4640
848:0006-3363
799:0268-1161
750:0268-1161
674:1058-0468
606:0021-8812
485:1747-3403
288:In humans
252:annexin V
248:apoptotic
212:viability
196:fertilize
136:annexin V
132:apoptosis
25:fertilize
1042:23337421
1001:23148085
856:16306419
807:19553239
758:18765400
692:23468098
614:15526798
575:23876533
534:23809504
493:17644989
455:23809499
411:41862891
403:26845061
343:See also
302:such as
200:motility
174:Z method
29:egg cell
949:3204150
905:3215823
683:3644127
81:swim-up
66:Methods
37:swim-up
1126:Family
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274:calves
244:YO-PRO
23:is to
1156:Semen
1060:(PDF)
407:S2CID
1038:PMID
997:PMID
989:ISSN
945:PMID
937:ISSN
901:PMID
893:ISSN
852:PMID
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803:PMID
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754:PMID
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688:PMID
670:ISSN
610:PMID
602:ISSN
571:PMID
530:PMID
489:PMID
481:ISSN
451:PMID
399:PMID
256:MACS
210:and
27:the
1028:doi
979:doi
975:145
929:doi
883:doi
834:doi
785:doi
736:doi
678:PMC
662:doi
594:doi
561:doi
557:100
520:doi
516:100
441:doi
437:100
391:doi
324:FDA
306:or
238:or
58:or
35:or
1122::
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957:^
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