William R. Boone

Improved cryopreserved semen fecundability in an alternating fresh-frozen artificial insemination program

This study with glycerol and increased number of motile sperm inseminated showed an increase in f to 10.4% for frozen semen and 27.4% for fresh semen. There is a significant (P less than 0.01) difference in pregnancy rate when greater than 100 million motile sperm are inseminated (31.3%) than when less than that number are inseminated (11.9%) in the first cycle when frozen semen is used.

Quality control in the in vitro fertilization laboratory.

Abstract Viability of gametes and embryos is explored as it relates to in vitro fertilization (IVF) and embryo culture (EC): exposure to cytotoxic materials, water quality, gas composition, temperature, hydrogen ion concentration and osmolality of media, and light exposure. The use of various assay techniques to strengthen quality control procedures in the IVF/EC laboratory is evaluated. In addition, original data concerning embryo toxicity of plasticware, changes in osmolality of culture medium over time, potential hazards of cleaning and sterilizing reusable labware and comparisons of 1-cell versus 2-cell embryo assays are discussed. Suggestions are then made concerning techniques to improve quality control in the IVF/EC laboratory.

Relative efficiency of therapeutic donor insemination using a luteinizing hormone monitor

A prospective randomized study was performed to evaluate the use of a urinary luteinizing hormone (LH) detection kit with 1 insemination as compared with 2 alternate day inseminations with timing based on previous cycle length and basal body temperature changes. The study involved 60 patients who underwent a total of 264 therapeutic donor insemination cycles using cryopreserved semen specimens. Patients alternated LH-kit timed cycles with cycles timed by non-LH methods for a total of 6 cycles or until pregnancy was achieved. Fecundability rates were 12.3% for LH-kit cycles and 5.3% for non-LH method cycles. The difference in outcome was not statistically significant. However, when the LH kit plus 1 insemination was compared with 2 inseminations timed by conventional methods, there appeared to be a distinct monetary and time expenditure advantage. These findings suggest that sufficient advantage may be derived from use of an LH kit to recommend its use on a routine basis for the timing of therapeutic donor insemination.

Freezing of mammalian embryos without the aid of a programmable freezer

A simple, inexpensive and repeatable method of freezing/thawing (F/T) mammalian concepti was developed with the use of 2-cell mouse embryos. Cryoprotectants, length of exposure to protectants, and subzero holding times before liquid nitrogen (LN2) exposure were examined in an effort to obtain an effective freezing protocol. Sequential examination of these variables provided data suggesting that 3.5 M dimethyl sulfoxide (DMSO) plus 0.25 M sucrose exposure for 5 minutes at room temperature, followed by a -30 degrees C environment for 90 minutes, best prepared embryos for LN2 storage. After thawing and culture, 48 of the 93 (52%) embryos developed to the blastocyst stage.

Relationship of heparin binding with computer-analyzed physical traits of human sperm**Supported by a University of Wisconsin biomedical research support grant (J.M.V.); a University of Wisconsin graduate school grant (J.M.V.); and a consortium grant from Atlantic Breeders Cooperative, NOBA, Sire Power, Inc., and Twenty-First Century Genetics (R.L.A.).

The objective of this study was to determine whether heparin affinity or the concentration of binding sites was related to sperm traits analyzed by a routine computerized semen analysis in human males requesting infertility evaluations. Saturation of heparin binding sites on sperm was achieved in 36 of 50 samples analyzed so that accurate estimates of dissociation constants (Kd) and binding site concentrations could be made. A broad range in Kds (18.2 to 284.5 nM/micrograms deoxyribonucleic acid [DNA]) and binding site concentrations (2.70 to 44.77 nM/micrograms DNA) was found. The binding affinity of sperm for heparin was significantly correlated with concentrations of cells in the ejaculate (r = 0.56), concentrations of motile sperm (r = 0.51), percentage of motile sperm (r = 0.33), and total numbers of ejaculated sperm (r = 0.37). The concentration of heparin binding sites also was correlated with concentration of cells in the ejaculate (r = 0.60), concentration of motile sperm (r = 0.50), percentage of motile sperm (r = 0.40), and total number of ejaculated sperm (r = 0.49).