Ooplasmic injection of elongating spermatids for the treatment of non- obstructive azoospermia

Sofikitis, Nikolaos; Yamamoto, Yasuhisa; Miyagawa, Ikuo; Mekras, G; Mio, Y.; Toda, Toshiko; Antypas, S.; Kawamura, Hiroshi; Kanakas, N.; Antoniou, Nikolaos; Loutradis, Dimitrios; Mantzavinos, Themis; Kalianidis, K; Agapitos, E.

doi:10.1093/humrep/13.3.709

Cited by 51 publications

(44 citation statements)

References 34 publications

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“…After optimization of the isolation procedure, the focus was on finding the optimal conditions for culturing the cells. The medium constituents were chosen from a literature review of previously published data on factors affecting viability of germ cells (Jutte et al, 1981; Caussanel et al, 1996; Sofikitis et al, 1998; Pentikäinen et al, 2000; Tesarik et al, 2002; Peterson and Soder, 2006; Oatley et al, 2007). For instance, lactate was added as the primary energy source for spermatids, along with factors secreted by Sertoli cells that have been shown to play an important role in maintaining viability of meiotic and post‐meiotic germ cells (Table 1).…”

Section: Resultsmentioning

confidence: 99%

An in vitro, short‐term culture method for mammalian haploid round spermatids amenable for molecular manipulation

Dehnugara

Dhar

2011

Molecular Reproduction Devel

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Extensive chromatin remodeling is a characteristic feature of mammalian spermiogenesis. To date, methods for the molecular manipulation of haploid spermatids are not available as there is a lack of a well-established culture system. Biochemical experiments and knockout studies reveal only the final outcome; studying the incremental details of the intricate mechanisms involved is still a challenge. We have established an in vitro culture system for pure haploid round spermatids isolated from rat testes that can be maintained with good viability for up to 72 hr. Changes in cell morphology and flagellar growth were also studied in the cultured spermatids. Further, we have demonstrated that upon treatment of cells with specific histone deacetylase inhibitors, sodium butyrate and trichostatin A, there is an increase in the hyperacetylation status of histone H4, mimicking an important event characteristic of histone replacement process that occurs during later stages of spermiogenesis. We have also tried various methods for introducing DNA and protein into these round spermatids in culture, and report that while DNA transfection is still a challenging task, protein transfection could be achieved using Chariot™ peptide as a transfection reagent. Thus, the method described here sets a stage to study the molecular roles of spermatid-specific proteins and chromatin remodelers in the cellular context.

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Section: Resultsmentioning

confidence: 99%

An in vitro, short‐term culture method for mammalian haploid round spermatids amenable for molecular manipulation

Dehnugara

Dhar

2011

Molecular Reproduction Devel

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“…More recently, it has also been shown that normal embryo development could be obtained from a sterile mouse strain after ROSI (Sasagawa et al, 1998). In addition, in the human, babies have been delivered from round and elongated spermatids (Fishel et al, 1995(Fishel et al, , 1997Mansour et al, 1996;Tesarik and Mendoza, 1996a;Tesarik et al, 1996b;Antinori., 1997;Araki et al, 1997;Vanderwalmen et al, 1997;Barak et al, 1998, Bernabeu et al, 1998Sofikitis et al, 1998), but the efficacy of the process is not yet established. Although the underlying mechanism of this technique is not fully understood, the fact that fertilization, implantation, and birth of healthy infants can occur after the injection of spermatids into oocytes has led us to use this technique for infertile couples in those cases in which the technique represented their only chance of pregnancy.…”

Section: Discussionmentioning

confidence: 99%

“…Intact round spermatid injection (ROSI) has resulted in the delivery of healthy infants (Tesarik and Mendoza, 1996a;Tesarik et al, 1996b;Vanderzwalmen et al, 1997;Barak et al, 1998;Bernabeu et al, 1998), as has treatment with elongated spermatid injection (ELSI) (Fishel et al, 1995(Fishel et al, , 1997Mansour et al, 1996;Antinori, 1997;Araki et al, 1997;Vanderzwalmen et al, 1997;Sofikitis et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Successful metaphase chromosome analysis of human elongated spermatids using mouse oocytes

Araki¹,

Ogawa²,

Ohno³

et al. 1999

Molecular Human Reproduction

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Human elongated spermatids from azoospermic patients were inserted into mouse oocytes by intracytoplasmic sperm injection (ICSI). The injection resulted in survival rates of 46.5% (180 out of 387) and activation rates of 36.1% (65 out of 180). The rate of two pronuclear (2PN) formation was 35.4% (23 out of 65). Only 34.8% (eight out of 23) metaphase chromosome spreads from 2PN zygotes could be analysed; however, all were of normal karyotype. Cytogenetic analysis at the first metaphase revealed that human elongated spermatid chromosomes were able to undergo replication in a heterogeneous environment.

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“…One might consider using iPS cell-derived spermatids in the clinical setting. However, although oocytes have been fertilized with elongated spermatids [39,40], they were insufficiently fertilized with premature, round spermatids, resulting in poor embryonic development [41,42,43]. Successful fertilization of oocytes with more matured male germ cells in vitro needs to be examined in preclinical research.…”

Section: The Induction Of Germ Cells From Ips Cellsmentioning

confidence: 99%

Human iPS Cell-Derived Germ Cells: Current Status and Clinical Potential

Ishii

2014

JCM

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Recently, fertile spermatozoa and oocytes were generated from mouse induced pluripotent (iPS) cells using a combined in vitro and in vivo induction system. With regard to germ cell induction from human iPS cells, progress has been made particularly in the male germline, demonstrating in vitro generation of haploid, round spermatids. Although iPS-derived germ cells are expected to be developed to yield a form of assisted reproductive technology (ART) that can address unmet reproductive needs, genetic and/or epigenetic instabilities abound in iPS cell generation and germ cell induction. In addition, there is still room to improve the induction protocol in the female germline. However, rapid advances in stem cell research are likely to make such obstacles surmountable, potentially translating induced germ cells into the clinical setting in the immediate future. This review examines the current status of the induction of germ cells from human iPS cells and discusses the clinical potential, as well as future directions.

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Ooplasmic injection of elongating spermatids for the treatment of non- obstructive azoospermia

Cited by 51 publications

References 34 publications

An in vitro, short‐term culture method for mammalian haploid round spermatids amenable for molecular manipulation

An in vitro, short‐term culture method for mammalian haploid round spermatids amenable for molecular manipulation

Successful metaphase chromosome analysis of human elongated spermatids using mouse oocytes

Human iPS Cell-Derived Germ Cells: Current Status and Clinical Potential

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