Sex preselection by flow cytometric separation of X and Y chromosome-bearing sperm based on DNA difference: a review

1995 ◽  
Vol 7 (4) ◽  
pp. 893 ◽  
Author(s):  
LA Johnson
Keyword(s):  
Y Chromosome ◽  
Sex Chromosome ◽  
Human Sperm ◽  
Specific Marker ◽  
Current Form ◽  
Vitro Fertilization ◽  
Flow Cytometric ◽  
Flow Cytometric Sorting

Recent research on the flow cytometry of sperm for the purpose of predetermining gender of offspring has led to a validated method to separate X from Y chromosome-bearing sperm for use with in vitro fertilization and embryo transfer, intratubal insemination or intracytoplasmic sperm injection. The basis for the method is the sex chromosome-specific marker, DNA, which is present in greater amounts in X-bearing sperm than in Y-bearing sperm of mammals. Sperm are exposed to the vital dye Hoechst 33342 which binds to the minor groove of the DNA helix. Flow cytometric sorting of the sperm using a laser as the excitation source results in populations of Y- or X-bearing sperm that are 85-90% pure. Several hundred offspring have been produced from swine, rabbits, sheep and cattle that confirm the predicted sex. The method is currently being applied to the commercial embryo market. The method is not likely to be used in conjunction with standard cattle or swine artificial insemination practice in its current form since only about 4 x 10(5) sorted sperm can be produced per hour of sorting. The technology has also been applied to human sperm for use by couples that are at risk to sex-linked disease expression in their offspring. Populations of human sperm have been sorted with X and Y purities of about 80% as confirmed by DNA probe technology and fluorescence in situ hybridization.

The relationship between membrane status and fertility of boar spermatozoa after flow cytometric sorting in the presence or absence of seminal plasma

1998 ◽  
Vol 10 (5) ◽  
pp. 433 ◽  
Author(s):  
W. M. C. Maxwell ◽  
C. R. Long ◽  
L. A. Johnson ◽  
J. R. Dobrinsky ◽  
G. R. Welch
Keyword(s):  
Seminal Plasma ◽  
Hoechst 33342 ◽  
Cell Sorter ◽  
Vitro Fertilization ◽  
Flow Cytometric ◽  
Collection Medium ◽  
Flow Cytometric Sorting ◽  
Boar Spermatozoa ◽  
The Relationship

The motility, viability (percent live), capacitation status and in vitro fertility of boar spermatozoa were examined, after staining with Hoechst 33342 and flow cytometric sorting in the absence or presence of seminal plasma. Viability was higher in unstained controls and when seminal plasma was present in the medium used to collect spermatozoa from the cell sorter than when seminal plasma was absent or in the staining extender only, but motility was highest when seminal plasma was included in the extender only, compared with the controls and other treatments. The proportions of capacitated spermatozoa were increased by sorting, but were lower when seminal plasma was present, rather than absent, from the staining extender and the collection medium. Compared with unstained controls, extension and staining without sorting only increased the proportion of capacitated spermatozoa after washing in preparation for in vitro fertilization. The percentages of polyspermic, penetrated and cleaved oocytes were lower when inseminated with unsorted (stained) than control (unstained) spermatozoa, regardless of the presence or absence of seminal plasma. These parameters were higher for sorted than for control spermatozoa in the absence of seminal plasma, but in its presence penetration and cleavage were substantially lower. The proportions of capacitated spermatozoa were lower when seminal plasma was present in the collection medium only than in the staining extender or when it was absent altogether, but the former treatment substantially reduced the proportions of polyspermic, penetrated and cleaved oocytes, and the proportion of blastocysts. These findings indicate that sperm capacitation associated with flow cytometric sorting can be reduced by the inclusion of seminal plasma in the collection medium, but this treatment reduces the ability of spermatozoa to fertilize oocytes in vitro under these conditions.

The effect of Y-chromosome alpha-satellite array length on the rate of sex chromosome disomy in human sperm

1996 ◽  
Vol 97 (6) ◽  
pp. 819-823 ◽  
Author(s):  
Michael A. Abruzzo ◽  
Darren K. Griffin ◽  
Elise A. Millie ◽  
Leon A. Sheean ◽  
Terry J. Hassold
Keyword(s):  
Y Chromosome ◽  
Sex Chromosome ◽  
Human Sperm ◽  
Alpha Satellite

Quality control in an in-vitro fertilization laboratory: use of human sperm survival studies

1989 ◽  
Vol 4 (5) ◽  
pp. 545-549 ◽  
Author(s):  
J. Diane Critchlow ◽  
Phillip L. Matson ◽  
C. Newman Maureen ◽  
Gregory Horne ◽  
Stephen A. Troup ◽  
...  
Keyword(s):  
Quality Control ◽  
In Vitro Fertilization ◽  
Human Sperm ◽  
Vitro Fertilization ◽  
Sperm Survival ◽  
Survival Studies

Temporal changes in the expression of the insulin-like growth factor II gene associated with tissue maturation in the human fetus

Development ◽  
1989 ◽  
Vol 106 (3) ◽  
pp. 543-554 ◽  
Author(s):  
A.L. Brice ◽  
J.E. Cheetham ◽  
V.N. Bolton ◽  
N.C. Hill ◽  
P.N. Schofield
Keyword(s):  
Growth Factor ◽  
Cell Types ◽  
Specific Cell ◽  
Insulin Like Growth Factor ◽  
Vitro Fertilization ◽  
Age Related ◽  
Factor Ii ◽  
Metanephric Blastema

The insulin-like growth factors are broadly distributed in the human conceptus and are thought to play a role in the growth and differentiation of tissues during development. Using in situ hybridization we have shown that a wide variety of specific cell types within tissues express the gene for insulin-like growth factor II at times of development from 18 days to 14 weeks of gestation. Examination of blastocysts produced by in vitro fertilization showed no expression, thus bracketing the time of first accumulation of IGF-II mRNA to between 5 and 18 days postfertilization. The pattern of IGF-II expression shows specific age-related differences in different tissues. In the kidney, for example, expression is found in the cells of the metanephric blastema which is dramatically reduced as the blastema differentiates. The reverse is also seen, and we have noted an increase in expression of IGF-II in the cytotrophoblast layer of the placenta with gestational age. The sites of expression do not correlate with areas of either high mitotic activity or specific types of differentiation, but the observed pattern of expression in the kidney, adrenal glands and liver suggests an explanation for the abnormally high IGF-II mRNA expression in developmental tumours such as Wilms' tumour.

Prematurely condensed human sperm chromosomes after in vitro fertilization (IVF)

1986 ◽  
Vol 74 (4) ◽  
pp. 441-443 ◽  
Author(s):  
H. Schmiady ◽  
K. Sperling ◽  
H. Kentenich ◽  
M. Stauber
Keyword(s):  
In Vitro Fertilization ◽  
Human Sperm ◽  
Vitro Fertilization ◽  
Sperm Chromosomes ◽  
Human Sperm Chromosomes

Development and implementation of intracytoplasmic sperm injection (ICSI)

1995 ◽  
Vol 7 (2) ◽  
pp. 211 ◽  
Author(s):  
GD Palermo ◽  
J Cohen ◽  
M Alikani ◽  
A Adler ◽  
Z Rosenwaks
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Human Sperm ◽  
Fertilization Rate ◽  
Semen Parameters ◽  
Ongoing Pregnancy Rate ◽  
Vitro Fertilization ◽  
Human Oocytes ◽  
Intracytoplasmic Injection ◽  
Previously Treated

The purpose of this paper is to elucidate the experimental steps that led to the development of intracytoplasmic sperm injection (ICSI) and its application in the human. ICSI has become the most successful micromanipulation procedure for treating male infertility. A total of 355 in vitro fertilization (IVF) cycles utilizing ICSI are described; 180 couples were previously treated in 509 IVF cycles but achieved no fertilization and 175 couples could not be treated by IVF because of extremely poor semen parameters. Of the 3063 metaphase II (M II) oocytes retrieved, 2970 were injected with a survival rate of 93.6%, yielding 1917 bipronuclear zygotes (64.5%). In 148 patients, a foetal heart was evidenced by ultrasound; 11 of these patients miscarried between 7 and 13 weeks of gestation. The ongoing pregnancy rate was 38.6% (137/355) per retrieval and 40.5% (137/338) per embryo replacement. At the time of writing, there were 22 deliveries and one therapeutic abortion for a trisomy 21 chromosomal abnormality. In addition, 66 singleton, 37 twin, 10 triplet and 1 quadruplet pregnancies were ongoing. The concentration of motile spermatozoa in the ejaculate only slightly influenced the fertilization rate (P < 0.001) and the pregnancy outcome (P < 0.01). A preliminary injection procedure utilizing intracytoplasmic injection of isolated sperm heads was performed in 35 M II human oocytes with resultant fertilization and cleavage rates of 74% and 73% respectively. Skills in ICSI were acquired by injecting hamster and unfertilized human oocytes with human sperm. ICSI can be used to successfully treat couples who have failed IVF or who have too few spermatozoa for conventional in vitro insemination.(ABSTRACT TRUNCATED AT 250 WORDS)

Recurrent In Vitro Fertilization Failure Evaluated by Fluorescence In Situ Hybridization: A Case Report 1

1998 ◽  
Vol 69 (3) ◽  
pp. 558-560 ◽  
Author(s):  
M Grossmann M.S. ◽  
J.M Calafell Ph.D. ◽  
V Moreno Ph.D., ◽  
J Balasch M.D., ◽  
J.A Vanrell M.D., ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Case Report ◽  
In Vitro Fertilization ◽  
Fluorescence In Situ Hybridization ◽  
Fertilization Failure ◽  
Vitro Fertilization
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