Evaluation of the TMRW vapor phase cryostorage platform using reproductive specimens and in vitro extended human embryo culture.

OBJECTIVE: To assess the impact of shipment and storage of sperm, oocytes, and blastocysts in vapor phase nitrogen compared with static storage in liquid phase nitrogen. DESIGN: Prospective cohort-matched study. SETTING: Multiple in vitro fertilization laboratories in an in vitro fertilization network. PATIENT(S): Fifty-eight human embryos, 32 human oocytes, 15 units of bovine semen. INTERVENTION(S): Vapor vs. liquid nitrogen. MAIN OUTCOME MEASURE(S): The postwarming survival of oocytes, sperm, and blastocysts, and the developmental potential of blastocysts during in vitro extended culture. RESULT(S): Custom-designed labware, for use with the TMRW platform, enables continuous temperature monitoring during shipment and/or storage in the vapor phase robotic storage system. The highest temperature recorded for specimens shipped to a domestic laboratory was -180.2 °C with a mean ± SD of -190.4 ± 0.5 °C during shipment and -181.1 ± 0.6 °C during storage. Likewise, specimens shipped internationally had a high of -180.2 °C with a mean ± SD of -193.5 ± 0.6 °C during shipment and -181.2 ± 0.7 °C during storage. Results from the extended culture assays have revealed no deleterious effect of shipment and storage in nitrogen vapor. The viability of mammalian gametes and embryos was equivalent between the vapor phase and liquid phase storage. CONCLUSION(S): The evaluated system did not have any deleterious effects on the postwarming survival of sperm, oocytes, and blastocysts. The postwarming developmental potential of human blastocysts during in vitro extended culture was unaffected by storage and handling in the vapor phase nitrogen TMRW platform when compared with static liquid phase nitrogen storage. Our results suggest that the vapor phase cryostorage platform is a safe system to handle and store reproductive specimens for human assisted reproductive technology.

Culture of Human Preimplantation Embryos in a Clinical ART Setting.

In vitro fertilization (IVF) and embryo culture in the human is a unique endeavor. Human assisted reproductive technology (ART) is practiced clinically to treat infertility. Due to the obvious ethical considerations of ART as applied to human medicine, only rarely is embryo culture undertaken for research purposes. As most IVF clinics are for profit businesses, a robust industry has developed to supply embryologists with quality control-tested media, equipment, and supplies necessary to support human IVF laboratory operation. Moreover, commercial items are preferred for efficiency and consistency, and strict quality control is required by accrediting organizations. As such, very little manual formulation or preparation of culture medium is typically required. Although human embryo culture is performed clinically, there is a surprising degree of variability in the details of the techniques used. In this chapter, we describe state-of-the-art procedures for gamete collection, in vitro fertilization, embryo culture, and embryo transfer that result in excellent blastocyst development and pregnancy rates for patients seeking treatment for infertility.

First successful delivery in Texas using vitrified human oocytes: a case report.

BACKGROUND: Successful pregnancies from cryopreserved oocytes are rare, but oocyte vitrification holds great promise for women in need of preserving their fertility due to illnesses that require treatments such as chemotherapy or radiation which cause irreversible depletion of ovarian reserve. The technique may also eventually be beneficial to women who wish to delay pregnancy to pursue educational and professional goals. Attempts at oocyte cryopreservation have until recently been quite disappointing due to three main problems: (1) high water content and intracellular ice crystal formation upon freezing and subsequent meiotic spindle damage, (2) zona pellucida hardening during cryopreservation and thus difficulty with subsequent fertilization and (3) the relatively large size of the cell and thus an unfavorable surface-to-volume ratio for equilibrium of solutes. These roadblocks have been gradually overcome by the use of improved cryoprotectants, intracytoplasmic sperm injection for fertilization, and the replacement of sodium in freezing media with an osmolyte. The net effect has been a substantial increase in oocyte survival and viability after cryopreservation. CASE: We report the first live births in Texas using vitrified oocytes. CONCLUSION: Vitrification may serve as a useful tool in the preservation of oocytes for women who wish to delay child bearing for medical or social reasons.

Excellent embryo quality obtained from vitrified oocytes.

Oocyte vitrification has the potential to expand tremendously the reproductive options for women desiring fertility preservation. Indeed, the number of publications regarding the reliability and efficiency of oocyte vitrification has increased exponentially over the past decade. The application of the technique to standard reproductive technologies depends on not only oocyte survival, but also the developmental potential and clinical outcomes from the resultant embryos. The aim of this study, therefore, was to examine the embryo quality obtained from vitrified, warmed oocytes in a standard infertility population. The results demonstrate that high quality blastocyst development can be expected with subsequent excellent obstetric and perinatal outcomes. These data support oocyte vitrification as a promising technique for fertility preservation.