Validation-verification of a highly effective, practical human testicular tissue in vitro culture-cryopreservation procedure aimed to optimize pre-freeze and post-thaw motility.

Purpose: The aim of our paper was to validate a testicular biopsy procedure that simplifies handling, processing, and cryopreservation, while at the same time optimizes sperm motility before freezing and after thawing.

Methods: Two prospective studies were conducted to verify, optimize, and understand the virtues of pre-freeze testicular tissue IVC at different temperatures (21, 30, or 37 °C). Testicular tissue was obtained from clinical specimens designated for whole tissue cryopreservation (i.

Frequency of postinsemination infections as reported by donor semen recipients.

Over an 18-year period, 47 women claimed to have acquired an infection after insemination with donor semen. This translates to an infection rate of less than 1 per 10,000 inseminations.

Evidence for stem cells in cultures of mouse prostate epithelial cells.

Background: Primary cultures of mouse prostatic epithelial cells were studied to identify cells having stem cell characteristics.

Methods: Culture conditions supporting the growth of mouse prostatic epithelial cells were identified. Immunostaining identified cells expressing cytokeratin 5 (K5), cytokeratin 8 (K8), and bcl-2.

Discoordinate regulation of different K channels in cultured rat skeletal muscle by nerve growth factor.

We investigated the effects of nerve growth factor (NGF) on expression of K+ channels in cultured skeletal muscle. The channels studied were (1) charybdotoxin (ChTx)-sensitive channels by using a polyclonal antibody raised in rabbits against ChTx, (2) Kv1.5 voltage-sensitive channels, and (3) apamin-sensitive (afterhyperpolarization) channels.

Calcium concentration and fertilization by subzonal insemination of a single spermatozoon in mouse oocytes.

The effect of calcium concentration in culture medium on the fertilization of subzonally microinseminated mouse oocytes was examined. Oocytes were injected with a single spermatozoon so that the sperm head was forced to adhere onto the ooplasmic membrane with a micromanipulation technique. For the inseminations, epididymal spermatozoa preincubated in culture medium and those treated with ionophore A23187 were used.

Subzonal insemination of a single mouse spermatozoon with a personal computer-controlled micromanipulation system.

A personal computer-controlled micromanipulation system was developed for automatic injection of spermatozoa into the perivitelline space of mouse ova. A pair of three-dimensional hydraulic micromanipulators driven by pulse motors was used for this automatic system. The pulse signals that regulate the motors are initiated by the computer program, and these signals cause the micromanipulator to move the microtool precisely.

Subzonal insemination with a single spermatozoon using manipulation assisted sperm adhesion onto the ooplasmic membrane in mouse ova.

A simple and successful method of microinjection of a single spermatozoon under the zona pellucida of a mouse oocyte has been developed. A characteristic of this method is that the tip of the sperm injection needle pierces the zona pellucida without touching the ooplasmic membrane. All the ova (277) used for this series of experiments had normal morphology after the injection procedure.

Micromanipulation in a center for reproductive medicine.

Several variations of micromanipulation of the female gamete (zona drilling, zona cracking, ooplasmic sperm injection, partial zona dissection) have been applied recently to human IVF to overcome severe male factor. Of the first 16 cycles attempted using partial zona dissection, one pregnancy resulted leading to a normal term delivery. Careful removal of the coronal cells, as well as stepwise removal of sucrose postpartial zona dissection, will facilitate this procedure and greatly reduce potential damage to the oocyte by pH, mechanical, or thermal injury.

The effect of ultrasound coupling gels on sperm motility in vitro.

Several of the most commonly used ultrasound coupling gels were found to adversely affect sperm motility. These gels should not be used in patients undergoing insemination immediately after a transvaginal pelvic ultrasonographic examination. Instead, we recommend the use of Proception Sperm Nutrient Douche, Nutrient Mixture F-10 (HAM), or nonlubrication of the condom-covered transducer.

Measurement of leukocyte motility and chemotaxis parameters with a linear under-agarose migration assay.

The interpretation of quantitative assays for leukocyte chemotactic migration is usually made in terms of measurements such as leading front distance, total migrating cells, and leukotactic index. These quantities allow comparison of cellular migration behavior under specified conditions. They are not useful; however, for comparisons between systems or for correlation with in vivo performance, because they depend upon specific physical aspects of the assay system, such as the geometry, chemoattractant concentration and diffusivity, and observation time.

Analysis of the linear under-agarose leukocyte chemotaxis assay.

Random and chemotactic movement of leukocytes appear to be key processes in the host inflammatory response. Abnormalities in leukocyte motility and chemosensory behavior have been implicated in a large number of pathological conditions, but there is no adequate quantitative understanding of these properties. In this paper we present an approach for determination of phenomenological cell motility and chemotaxis parameters, by analysis of a common leukocyte migration assay.

The recurrent varicocele--a poorly recognized problem.

A varicocele is most likely the result of an incompetent vein with retrograde flow of blood into the scrotum. Surgical ligation of this incompetent system is the present-day procedure of choice. In our series of 225 varicocelectomies, there have been three definite recurrences, due to either recollateralization or failure to ligate all branches of this venous plexus.