The Experts below are selected from a list of 237 Experts worldwide ranked by ideXlab platform
L A Johnson - One of the best experts on this subject based on the ideXlab platform.
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microsort Sperm Sorting causes no increase in major malformation rate
Reproduction Fertility and Development, 2016Co-Authors: Donald P Marazzo, David Karabinus, L A Johnson, Joseph D SchulmanAbstract:The purpose of the present study was to evaluate the safety of MicroSort (MicroSort Division, GIVF, Fairfax, VA, USA) Sperm Sorting by monitoring major malformations in infants and fetuses conceived using sorted Spermatozoa. Data were collected in a prospective protocol with monitoring that began from conception through birth until 1 year of life. Comprehensive ascertainment identified fetuses and stillbirths with malformations after 16 weeks gestation, pregnancies terminated for malformations and babies with major malformations. Outcomes in MicroSort pregnancies were compared with outcomes in published studies that used active and comprehensive ascertainment of malformations in the general population and in pregnancies established after assisted reproduction. Using comprehensive outcomes from all pregnancies, the rate of major malformations in MicroSort pregnancies conceived after IVF with or without intracytoplasmic Sperm injection was 7.8%; this did not differ significantly from the rates reported in the three assisted reproductive technology control studies not associated with MicroSort (8.6%, 9.2% and 8.3%). Similarly, the rate of major malformations in MicroSort pregnancies initiated with intrauterine insemination was 6.0%, not significantly different from that reported in non-assisted reproductive technology pregnancies not associated with MicroSort (6.9%, 4.6% and 5.7%). Prospective record review of pregnancy outcomes and paediatric evaluation to 1 year indicate no association between MicroSort Sperm Sorting and major malformations.
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preselection of sex of offspring in swine for production current status of the process and its application
Theriogenology, 2005Co-Authors: L A Johnson, J M Vazquez, W M C Maxwell, Detlef Rath, J R DobrinskyAbstract:It is estimated that as many as 30,000 offspring, mostly cattle, have been produced in the past 5 years using AI or some other means of transport with Spermatozoa sexed by flow cytometric Sperm Sorting and DNA as the marker of differentiation. It is well documented that the only marker in Sperm that can be effectively used for the separation of X- and Y-chromosome bearing Spermatozoa is DNA. The method, as it is currently used worldwide, is commonly known as the Beltsville Sperm Sexing Technology. The method is based on the separation of Sperm using flow cytometric Sorting to sort fluorescently (Hoechst 33342) labeled Sperm based on their relative content of DNA within each population of X- and Y-Spermatozoa. Currently, Sperm can be produced routinely at a rate of 15 million X- and an equal number of Y-Sperm per hour. The technology is being applied in livestock, laboratory animals, and zoo animals; and in humans with a success rate of 90-95% in shifting the sex ratio of offspring. Delivery of sexed Sperm to the site of fertilization varies with species. Conventional AI, intrauterine insemination, intra-tubal insemination, IVF with embryo transfer and deep intrauterine insemination are effectively used to obtain pregnancies dependent on species. Although Sperm of all species can be sorted with high purity, achieving pregnancies with the low numbers of Sperm needed for commercial application remains particularly elusive in swine. Deep intrauterine insemination with 50-100 million sexed boar Sperm per AI has given encouragement to the view that insemination with one-fiftieth of the standard insemination number will be sufficient to achieve pregnancies with sexed Sperm when specialized catheters are used. Catheter design, volume of inseminate, number of sexed Sperm are areas where further development is needed before routine inseminations with sexed Sperm can be conducted in swine. Cryopreservation of sex-sorted Sperm has been routinely applied in cattle. Although piglets have been born from frozen sex-sorted boar Sperm, freezing and processing protocols in combination with sex-sorted Sperm are not yet optimal for routine use. This review will discuss the most recent results and advances in sex-Sorting swine Sperm with emphasis on what developments must take place for the sexing technology to be applied in commercial practice.
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flow cytometric Sperm Sorting effects of varying laser power on embryo development in swine
Molecular Reproduction and Development, 2002Co-Authors: L A Johnson, G R Welch, H D Guthrie, Wesley M Garrett, J R DobrinskyAbstract:This study was conducted to determine fertilization rate and embryo development using the Beltsville Sperm Sexing Technology with two different laser power outputs, 25 and 125 milliwatts (mW). Freshly ejaculated boar semen was diluted; one aliquot was not stained or sorted (nonsort) and a second aliquot was stained with Hoechst 33342 and sorted as a complete population, not separated into X and Y populations (all-sort). Ovulation controlled gilts were surgically inseminated with 2 10 5 Spermatozoa (44- 46 hr after human chorionic gonadotropin (hCG)) into the isthmus of each oviduct, one oviduct receiving nonsort and the other all-sort at 25 or 125 mW. A total of 426 embryos were flushed from oviducts at slaughter 43 hr after laparotomy and prepared for determination of fertilization and cleavage rates using confocal laser microscopy for analysis of actin cytoskeleton and chromatin configuration. The percentage of fertilized eggs and embryos was less for the 25 mW all-sort compared to nonsort or the 125 mW all-sort (77.9 vs. 96.3 and 96.2%, P 0.05) among treatments. However, the rate of embryo development was slower (P 0.05) from the nonsort or 25 mW all- sort. We conclude that the use of 125 mW laser power for Sorting boar Spermatozoa is advantageous to maintain high resolution separation and has no detrimental effect on embryo development compared to 25 mW. Mol. Reprod. Dev. 61: 87-92, 2002. fl 2002 Wiley-Liss, Inc.
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physiology of Spermatozoa at high dilution rates the influence of seminal plasma
Theriogenology, 1999Co-Authors: W M C Maxwell, L A JohnsonAbstract:Extensive dilution of Spermatozoa, as occurs during flow-cytometric Sperm Sorting, can reduce their motility and viability. These effects may be minimized by the use of appropriate dilution and collection media, containing balanced salts, energy sources, egg yolk and some protein. Dilution and flow-cytometric Sorting of Spermatozoa, which involves the removal of seminal plasma, also destabilizes Sperm membranes leading to functional capacitation. This membrane destabilization renders the Spermatozoa immediately capable of fertilization in vitro, or in vivo after deposition close to the site of fertilization, but shortens their lifespan, resulting in premature death if the cells are deposited in the female tract distant from the site of fertilization or are held in vitro at standard storage temperatures. This functional capacitation can be reversed in boar Spermatozoa by inclusion of seminal plasma in the medium used to collect the cells from the cell sorter and, consequently, reduces their in vitro fertility. It has yet to be determined whether seminal plasma would have similar effects on flow cytometrically sorted Spermatozoa of other species, and what its effects might be on the in vivo fertility of flow sorted boar Spermatozoa.
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the beltsville Sperm sexing technology high speed Sperm Sorting gives improved Sperm output for in vitro fertilization and ai
Journal of Animal Science, 1999Co-Authors: L A Johnson, G R Welch, Wim RensAbstract:The Beltsville Sperm sexing technology is currently the only effective means of altering the sex ratio of offspring in livestock. The method is based on the flow-cytometric separation of X- and Y-chromosome-bearing Sperm based on X/Y DNA content difference. It is an effective means of producing progeny of predetermined sex in cattle, swine, sheep, and laboratory animals. The method involves treating Sperm with a DNA-binding fluorochrome, Hoechst 33342, and flow-cytometrically Sorting them into separate X and Y populations that can subsequently be used for surgical intratubal or intrauterine insemination, deep-uterine insemination, regular artificial insemination in some cases, in vitro fertilization to produce sexed embryos for transfer, and intracytoplasmic Sperm injection of ova. Skewed sex ratios of 85 to 95% of one sex or the other have been repeatably achieved in most species. The method has been used worldwide to produce several hundred morphologically normal animal offspring of the predicted sex. It has also been validated in the laboratory using DNA reanalysis of the sorted Sperm populations and by fluorescence in situ hybridization and PCR of individual Sperm. We developed a new orienting nozzle that we have fitted to both conventional and high-speed cell sorters that have been modified for Sperm Sorting. Recently we completed the adaptation of the new orienting nozzle to a Cytomation MoFlo high-speed cell sorter modified for Sperm. This adaptation of the nozzle has increased the overall production rate of sorted X and Y Sperm from about .35 million/h to 5 or 6 million Sperm/h (each population). Calves have been born from cows artificially inseminated using conventional technique and sexed Sperm. In addition, numerous litters of pigs have been born after transfer of embryos produced from X or Y sorted Sperm.
Fangang Tseng - One of the best experts on this subject based on the ideXlab platform.
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high throughput flowing upstream Sperm Sorting in a retarding flow field for human semen analysis
Analyst, 2017Co-Authors: Pengchun Chen, Yunan Lin, Lichern Pan, Fangang Tseng, Chia Woei WangAbstract:In this paper, we propose a microfluidic device capable of generating a retarding flow field for the Sorting and separation of human motile Sperm in a high-throughput manner. The proposed Sorting/separation process begins with a rapid flow field in a straight-flow zone to carry Sperm into a Sorting zone to maintain the Sperm’s mobility. The Sorting zone consists of a diffuser-type Sperm sorter to differentiate Sperm with different motilities based on the flowing upstream nature of human Sperm in a retarding flow field. The dead Sperm will then be separated from the live ones by passing through a dumbbell flow field to the outlet for disposal. The proposed flowing upstream Sperm sorter (FUSS) is designed to imitate the selection mechanism found in the female body when Sperm swim into the uterus. The experimental results demonstrate the utility of this device with regard to throughput (approximately 200 000 Sperm per minute and a maximum of 200 million cells per mL), efficiency (90% of selected Sperm are mobile), and the ability to select Sperm with high motility (∼20% of Sperm with a velocity exceeding 120 μm s−1). The proposed device is suitable for intrauterine insemination as well as in vitro fertilization thanks to the highly efficient Sorting process not interfering with the natural function and energy resource of human Sperm.
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high throughput Sperm Sorting in a micro diffuser type fluidic system
International Conference on Micro Electro Mechanical Systems, 2013Co-Authors: Yunan Lin, Pengchun Chen, Lichern Pan, Fangang TsengAbstract:In this study, we propose a microfluidic device with a diffuser type chamber to differentiate the Sperm velocity by gradually lowering down the flow speed in diffuser by gradually increasing the width of the channel. When the Sperms' moving speed match the flow speed, the Sperms tend to have the ability to flow against the stream, thus separating the Sperms at different specific positions along the diffuser according to their motility becomes gentle and fast. Most of the motile Sperm will stay in the expanding area and the non-motile Sperms will drift to outlet.
Joerg Lahann - One of the best experts on this subject based on the ideXlab platform.
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A surface-modified Sperm Sorting device with long-term stability
Biomedical Microdevices, 2006Co-Authors: Jason M. Wu, Yaokuang Chung, Kimberly J. Belford, Gary D Smith, Shuichi Takayama, Joerg LahannAbstract:Microfluidic devices fabricated from poly (dime- thylsiloxane) (PDMS) offer the ability to improve our biological and medical capabilities. Although PDMS offers a range of intriguing benefits for biomedical applications, the intrinsically hydrophobic nature of PDMS may impede with the tremendous potential of these devices. Here, we describe a PDMS-based Sperm Sorting device, which has been surface-modified via graft-co-polymerization of poly(ethylene glycol) methyl ether methacrylate to create a moderately hydrophilic and non-fouling surface. This process involves the exposure of PDMS to UV/ozone, which activates the PDMS surface to bond to the substrate and, at the same time, initiates the graft-co-polymerization from the PDMS surface. In this study, we confirmed long-term stability of surface-modified PDMS for up to 56 days based on Fourier transformation infrared spectroscopy (FTIR), contact angle measurements, and protein adsorption studies. Moreover, the applicability of our method to PDMS-based Sperm Sorting devices was demonstrated by successfully Sorting human Sperm.
Keiji Naruse - One of the best experts on this subject based on the ideXlab platform.
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Mechanomedicine
Biophysical Reviews, 2018Co-Authors: Keiji NaruseAbstract:It has been a long time since the term mechanobiology became widely accepted, and broad research approaches, ranging from basic biology to medical research, have been conducted from the perspective of mechanobiology. Our group created the term “mechanomedicine” focusing on the field encompassing studies of the pathology and treatment of various diseases based on the knowledge obtained from mechanobiological studies and have promoted studies in this field. In the respiratory and cardiovascular systems, not only humoral factors but also physical factors such as contraction and expansion phenomena, and feedback from such phenomena to tissues and cells are important stimuli for maintaining homeostasis. Loss of homeostasis is considered to lead to pathological conditions. This review aims to provide an overview of mechanomedicine by introducing several mechanosensitive channels including one particular type of mechanosensor that we discovered in the cardiovascular system and by describing stretchable three-dimensional cell culture scaffolds using self-assembled peptides, a highly motile Sperm sorter using a Sperm Sorting technique based on microfluidic mechanics, and a device to promote the development of fertilized ova.
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screening of Sperm velocity by fluid mechanical characteristics of a cyclo olefin polymer microfluidic Sperm Sorting device
Reproductive Biomedicine Online, 2012Co-Authors: Koji Matsuura, Mami Takenami, Yuka Kuroda, Toru Hyakutake, Shinichiro Yanase, Keiji NaruseAbstract:The microfluidic Sperm-Sorting (MFSS) device is a promising advancement for assisted reproductive technology. Previously, poly(dimethylsiloxiane) and quartz MFSS devices were developed and used for intracytoplasmic Sperm injection. However, these disposable devices were not clinically suitable for assisted reproduction, so a cyclo-olefin polymer MFSS (COP-MFSS) device was developed. By micromachining, two microfluidic channels with different heights and widths (chip A: 0.3 x 0.5 mm; chip B: 0.1 x 0.6 mm) were prepared. Sorted Sperm concentrations were similar in both microfluidic channels. Linear-velocity distribution using the microfluidic channel of chip B was higher than that of chip A. Using confocal fluorescence microscopy, it was found that the highest number of motile Spermatozoa swam across the laminar flow at the bottom of the microfluidic channel. The time required to swim across the laminar flow was longer at the bottom and top of the microfluidic channels than in the middle because of the low fluid velocity. These results experimentally demonstrated that the width of microfluidic channels should be increased in the region of laminar flow from the semen inlet to the outlet for unsorted Spermatozoa to selectively recover Spermatozoa with high linear velocity.
J R Dobrinsky - One of the best experts on this subject based on the ideXlab platform.
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preselection of sex of offspring in swine for production current status of the process and its application
Theriogenology, 2005Co-Authors: L A Johnson, J M Vazquez, W M C Maxwell, Detlef Rath, J R DobrinskyAbstract:It is estimated that as many as 30,000 offspring, mostly cattle, have been produced in the past 5 years using AI or some other means of transport with Spermatozoa sexed by flow cytometric Sperm Sorting and DNA as the marker of differentiation. It is well documented that the only marker in Sperm that can be effectively used for the separation of X- and Y-chromosome bearing Spermatozoa is DNA. The method, as it is currently used worldwide, is commonly known as the Beltsville Sperm Sexing Technology. The method is based on the separation of Sperm using flow cytometric Sorting to sort fluorescently (Hoechst 33342) labeled Sperm based on their relative content of DNA within each population of X- and Y-Spermatozoa. Currently, Sperm can be produced routinely at a rate of 15 million X- and an equal number of Y-Sperm per hour. The technology is being applied in livestock, laboratory animals, and zoo animals; and in humans with a success rate of 90-95% in shifting the sex ratio of offspring. Delivery of sexed Sperm to the site of fertilization varies with species. Conventional AI, intrauterine insemination, intra-tubal insemination, IVF with embryo transfer and deep intrauterine insemination are effectively used to obtain pregnancies dependent on species. Although Sperm of all species can be sorted with high purity, achieving pregnancies with the low numbers of Sperm needed for commercial application remains particularly elusive in swine. Deep intrauterine insemination with 50-100 million sexed boar Sperm per AI has given encouragement to the view that insemination with one-fiftieth of the standard insemination number will be sufficient to achieve pregnancies with sexed Sperm when specialized catheters are used. Catheter design, volume of inseminate, number of sexed Sperm are areas where further development is needed before routine inseminations with sexed Sperm can be conducted in swine. Cryopreservation of sex-sorted Sperm has been routinely applied in cattle. Although piglets have been born from frozen sex-sorted boar Sperm, freezing and processing protocols in combination with sex-sorted Sperm are not yet optimal for routine use. This review will discuss the most recent results and advances in sex-Sorting swine Sperm with emphasis on what developments must take place for the sexing technology to be applied in commercial practice.
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flow cytometric Sperm Sorting effects of varying laser power on embryo development in swine
Molecular Reproduction and Development, 2002Co-Authors: L A Johnson, G R Welch, H D Guthrie, Wesley M Garrett, J R DobrinskyAbstract:This study was conducted to determine fertilization rate and embryo development using the Beltsville Sperm Sexing Technology with two different laser power outputs, 25 and 125 milliwatts (mW). Freshly ejaculated boar semen was diluted; one aliquot was not stained or sorted (nonsort) and a second aliquot was stained with Hoechst 33342 and sorted as a complete population, not separated into X and Y populations (all-sort). Ovulation controlled gilts were surgically inseminated with 2 10 5 Spermatozoa (44- 46 hr after human chorionic gonadotropin (hCG)) into the isthmus of each oviduct, one oviduct receiving nonsort and the other all-sort at 25 or 125 mW. A total of 426 embryos were flushed from oviducts at slaughter 43 hr after laparotomy and prepared for determination of fertilization and cleavage rates using confocal laser microscopy for analysis of actin cytoskeleton and chromatin configuration. The percentage of fertilized eggs and embryos was less for the 25 mW all-sort compared to nonsort or the 125 mW all-sort (77.9 vs. 96.3 and 96.2%, P 0.05) among treatments. However, the rate of embryo development was slower (P 0.05) from the nonsort or 25 mW all- sort. We conclude that the use of 125 mW laser power for Sorting boar Spermatozoa is advantageous to maintain high resolution separation and has no detrimental effect on embryo development compared to 25 mW. Mol. Reprod. Dev. 61: 87-92, 2002. fl 2002 Wiley-Liss, Inc.