scholarly journals Mice generated after round spermatid injection into haploid two-cell blastomeres

Cell Research ◽  
2011 ◽  
Vol 21 (5) ◽  
pp. 854-858 ◽  
Author(s):  
Hui Yang ◽  
Linyu Shi ◽  
Charlie Degui Chen ◽  
Jinsong Li
Keyword(s):  
Round Spermatid ◽  
Round Spermatid Injection

Is Round Spermatid Injection (ROSI) a Therapy for Male Infertility?: ROSI in the Rhesus Monkey is Unsuccessful

2000 ◽  
Vol 74 (3) ◽  
pp. S68 ◽  
Author(s):  
L Hewitson ◽  
C Martinovich ◽  
C Simerly ◽  
T Dominko ◽  
G Schatten
Keyword(s):  
Male Infertility ◽  
Rhesus Monkey ◽  
Round Spermatid ◽  
Round Spermatid Injection

scholarly journals Transfer at the blastocyst stage of embryos derived from testicular round spermatid injection

2002 ◽  
Vol 17 (3) ◽  
pp. 741-743 ◽  
Author(s):  
B. Urman ◽  
C. Alatas ◽  
S. Aksoy ◽  
R. Mercan ◽  
A. Nuhoglu ◽  
...  
Keyword(s):  
Round Spermatid ◽  
Blastocyst Stage ◽  
Round Spermatid Injection

Round spermatids stained with MitoTracker can be used to produce offspring more simply

Zygote ◽  
2005 ◽  
Vol 13 (1) ◽  
pp. 55-61 ◽  
Author(s):  
Takafusa Hikichi ◽  
Satoshi Kishigami ◽  
Nguyen Van Thuan ◽  
Hiroshi Ohta ◽  
Eiji Mizutani ◽  
...  
Keyword(s):  
Embryo Development ◽  
Intracytoplasmic Sperm Injection ◽  
Round Spermatid ◽  
Testicular Cells ◽  
Infertility Treatments ◽  
Developmental Capacity ◽  
20 Nm ◽  
Harmful Effects ◽  
Round Spermatid Injection ◽  
Selection Of

Although both intracytoplasmic sperm injection (ICSI) and round spermatid injection (ROSI) are used in infertility treatments, the rate of offspring achieved with ROSI is low compared with that achieved with ICSI. The difficulty in correctly selecting round spermatids from testicular cells is one of the causes of this phenomenon. We easily selected live round spermatids from testicular cells stained with 20 nM MitoTracker, which visualizes mitochondria without killing the cell. Using this method, we divided round spermatids into three groups based on the polarization of their mitochondria, and performed ROSI. The rate of successful offspring achieved with MitoTracker-stained ROSI was the same in all groups. This indicates that changes in the polarization of mitochondria in round spermatids are not directly related to the developmental capacity of subsequently fertilized embryos. Because this staining has no harmful effects on embryo development, the selection of spermatids by MitoTracker under a fluorescence microscope should be useful in research into and the treatment of infertility.

scholarly journals The employment of strontium to activate mouse oocytes: effects on spermatid-injection outcome

Reproduction ◽  
2006 ◽  
Vol 131 (2) ◽  
pp. 259-267 ◽  
Author(s):  
Jean Loren ◽  
Orly Lacham-Kaplan
Keyword(s):  
Birth Outcomes ◽  
Round Spermatid ◽  
Oocyte Activation ◽  
Embryo Viability ◽  
Blastocyst Development ◽  
Incubation Periods ◽  
Artificial Oocyte Activation ◽  
The Impact ◽  
Post Implantation ◽  
Round Spermatid Injection

The present research investigated the effects of various strontium concentrations, in combination with different incubation periods, on mouse parthenogentic oocyte activation and blastocyst development. The results for blastocyst development showed a trend indicating that 10 mM strontium for 3 h was the optimal strontium protocol. Ethanol, an agent that incites oocyte activation via a monotonic rise in calcium, was employed as a control. The outcome of blastocyst formation arising from parthenogenic ethanol activation was significantly less (P < 0.001) than that achieved by the optimal strontium protocol. To assess the impact of strontium oocyte activation on embryo viability following fertilization with immature germ cells, the protocol of 10 mM strontium for 3 h was applied to oocytes injected with round spermatids and then compared with other protocols. The results indicate that following round-spermatid injection the benefits derived from strontium artificial oocyte activation are evident during both pre- and post-implantation development. However, in order to adjust the protocol to the most effective round-spermatid injection in relation to the oocyte cell cycle, injection was done 1.5 h after strontium activation followed by another 1.5 h activation in strontium. The implementation of round-spermatid injection in combination with this oocyte-activation protocol led to live-birth outcomes not significantly different to those outcomes obtained by mature spermatozoa.

Harmful or Not: Trichostatin A treatment of embryos generated by ICSI or ROSI

2006 ◽  
Vol 1 (3) ◽  
pp. 376-385 ◽  
Author(s):  
Satoshi Kishigami ◽  
Hiroshi Ohta ◽  
Eiji Mizutani ◽  
Sayaka Wakayama ◽  
Hong-Thuy Bui ◽  
...  
Keyword(s):  
Assisted Reproductive Technologies ◽  
Trichostatin A ◽  
Reproductive Technologies ◽  
Round Spermatid ◽  
Developmental Arrest ◽  
Offspring Production ◽  
Deacetylase Inhibitor ◽  
Cloning Method ◽  
Round Spermatid Injection ◽  
Parthenogenetic Embryos

AbstractTrichostatin A (TSA), a histone deacetylase inhibitor, is a known teratogen causing malformations such as vertebral fusions when applied during the postimplantation period; TSA also causes developmental arrest when applied during the preimplantation period. Regardless of these hindrances, we have succeeded in the establishment of an efficient somatic cloning method for the mouse where reconstructed embryos are treated with TSA. To elucidate this apparent discrepancy, we treated fertilized mouse embryos generated either by intracytoplasmic sperm injection (ICSI) or round spermatid injection (ROSI) with 50 nM TSA for 20 h after fertilization as well as parthenogenetic embryos and found that TSA treatment inhibited the preimplantation development of ICSI embryos but not ROSI or parthenogenetic embryos. And, although we often observed hypomorphism following TSA treatment in embryos grown to full term produced by both ICSI (av. of body weight: 1.7 g vs. 1.5 g) and ROSI (1.6 g vs. 1.2 g), TSA treatment reduced the offspring production rate for ICSI from 57% to 34% but not for ROSI from 30% to 36%. Thus, these data indicate that the effects, harmful or not, of TSA treatment on embryonic development depend on their nuclear derivations. Also, the resulting hypomorphism after TSA treatment is a caveat for this procedure in current Assisted Reproductive Technologies.

scholarly journals Recent Advances in Elongated and Round Spermatid Injection

2021 ◽  
Vol 8 (4) ◽  
pp. 234-237
Author(s):  
Fatih Gökalp ◽  
Sadık Görür
Keyword(s):  
Round Spermatid ◽  
Recent Advances ◽  
Round Spermatid Injection

375 UNEXPECTED SEVERE ABNORMALITIES IN MOUSE ROSI OFFSPRING

2007 ◽  
Vol 19 (1) ◽  
pp. 303
Author(s):  
P. N. Moreira ◽  
R. Fernández-González ◽  
M. Pérez-Crespo ◽  
P. Bermejo ◽  
J. D. Hourcade ◽  
...  
Keyword(s):  
Testicular Tumor ◽  
Round Spermatid ◽  
Sperm Cells ◽  
Healthy Children ◽  
Clinical Value ◽  
Cell Stage ◽  
Infertile Men ◽  
Vitro Development ◽  
Round Spermatid Injection

Live offspring resulting from round spermatid injection (ROSI) was first accomplished in the mouse, but similar success has been obtained in rat, hamster, rabbit, mastomys, pig, monkey, and human. ROSI has received clinical attention because some infertile men have no spermatozoa or just a very few in their testes, and these are difficult to harvest and are frequently dead and deformed. Although some clinicians were able to generate healthy children by ROSI, others could not (reviewed in Yanagimachi 2004 Reprod. Biomed. Online 9). The clinical value of ROSI has been widely debated. It remains unclear if post-meiotic and pre-fertilization modifications of sperm cells are necessary to ensure normal development. In order to answer this question, we decided to study and compare mouse offspring generated by ROSI and intracytoplasmic sperm injection (ICSI). ROSI and ICSI with fresh sperm cells were carried out in the B6D2 mouse strain as described (Marh et al. 2003 Biol. Reprod. 69, 169–176; Moreira et al. 2005 Hum. Reprod. 20, 3313–3317). In vitro-produced embryos were transferred at the 2-cell stage into Day 1 pseudopregnant females. As shown in Table 1 oocyte survival after injection was significantly higher (z-test, P &lt; 0.05) with ICSI (91%) than with ROSI (68%). The proportion of live offspring obtained by ICSI was also significantly higher (26% vs. 6%; z-test, P &lt; 0.05). Moreover, fertilization with spermatozoa produced healthy offspring more efficiently than with round spermatids. Out of 30 live offspring generated by ROSI, 6 (20%) presented severe abnormalities during their first 6–8 weeks of age. One ROSI animal presented an abnormally swollen skull (hydroencephaly) with a very thin and soft cranial wall. Another developed a subcutaneous engrossment of the forehead, producing a crest-like appearance. Three others presented deviations in their vertebral columns (hyperkyphosis and scoliosis), and recently a testicular tumor was detected in another animal. These types of malformations were not observed in the control offspring. In our experience, very rarely are they observed after ICSI or in naturally mated animals. To our knowledge, and although the risks of the ROSI procedure have been extensively highlighted in human and other species, the phenotypic abnormalities observed in this study have never been reported. Presently, we keep monitoring these animals as they age, as part of an ambitious plan that is also intended to characterize and understand the origin of the possible phenotypic consequences of the ROSI procedure. Table 1. In vitro development and development to term of B6D2 mouse embryos generated by ROSI or ICSI

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