Fourteen babies born after round spermatid injection into human oocytes

During the human in vitro fertilization procedure in the assisted reproductive technology, intracytoplasmic sperm injection is routinely used to inject a spermatozoon or a less mature elongating spermatid into the oocyte. In some infertile men, round spermatids (haploid male germ cells that have completed meiosis) are the most mature cells visible during testicular biopsy. The microsurgical injection of a round spermatid into an oocyte as a substitute is commonly referred to as round spermatid injection (ROSI). Currently, human ROSI is considered a very inefficient procedure and of no clinical value. Herein, we report the birth and development of 14 children born to 12 women following ROSI of 734 oocytes previously activated by an electric current. The round spermatids came from men who had been diagnosed as not having spermatozoa or elongated spermatids by andrologists at other hospitals after a first Micro-TESE. A key to our success was our ability to identify round spermatids accurately before oocyte injection. As of today, all children born after ROSI in our clinic are without any unusual physical, mental, or epigenetic problems. Thus, for men whose germ cells are unable to develop beyond the round spermatid stage, ROSI can, as a last resort, enable them to have their own genetic offspring.

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375 UNEXPECTED SEVERE ABNORMALITIES IN MOUSE ROSI OFFSPRING

Reproduction Fertility and Development ◽

10.1071/rdv19n1ab375 ◽

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 < 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 < 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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Advanced bioengineering of male germ stem cells to preserve fertility

Journal of Tissue Engineering ◽

10.1177/20417314211060590 ◽

2021 ◽

Vol 12 ◽

pp. 204173142110605

Author(s):

Hossein Eyni ◽

Sadegh Ghorbani ◽

Hojjatollah Nazari ◽

Marziyeh Hajialyani ◽

Sajad Razavi Bazaz ◽

...

Keyword(s):

Fertility Preservation ◽

Germ Cells ◽

Male Fertility ◽

Round Spermatid ◽

The Body ◽

Testicular Sperm Extraction ◽

Male Germ Cells ◽

Vitro Fertilization ◽

In Vitro Spermatogenesis

In modern life, several factors such as genetics, exposure to toxins, and aging have resulted in significant levels of male infertility, estimated to be approximately 18% worldwide. In response, substantial progress has been made to improve in vitro fertilization treatments (e.g. microsurgical testicular sperm extraction (m-TESE), intra-cytoplasmic sperm injection (ICSI), and round spermatid injection (ROSI)). Mimicking the structure of testicular natural extracellular matrices (ECM) outside of the body is one clear route toward complete in vitro spermatogenesis and male fertility preservation. Here, a new wave of technological innovations is underway applying regenerative medicine strategies to cell-tissue culture on natural or synthetic scaffolds supplemented with bioactive factors. The emergence of advanced bioengineered systems suggests new hope for male fertility preservation through development of functional male germ cells. To date, few studies aimed at in vitro spermatogenesis have resulted in relevant numbers of mature gametes. However, a substantial body of knowledge on conditions that are required to maintain and mature male germ cells in vitro is now in place. This review focuses on advanced bioengineering methods such as microfluidic systems, bio-fabricated scaffolds, and 3D organ culture applied to the germline for fertility preservation through in vitro spermatogenesis.

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