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PubMed2004

Evaluation of chromatin integrity in human sperm using acridine orange staining with different fixatives and after cryopreservation.

Staining of cells with acridine orange (AO) has been widely accepted as a predictor of DNA damage in many cell types. Because of variability of protocols used in previous studies, the AO staining technique has not been widely accepted as a screening test to predict DNA damage in human sperm. In order to further validate the use of AO staining, sperm were evaluated using numerous variations in the staining protocol. This study also elucidated the effects of cryopreservation on sperm DNA. Sperm fixation in Carnoy's solution showed significantly (P < 0.05) more DNA damage (29.9 +/- 4.5%) than 2% glutaraldehyde (14.4 +/- 2.1%), 4% paraformaldehyde (5.5 +/- 1.7%), no fixation (15.8 +/- 4.3%) but did not differ from Diff Quik solution (19.2 +/- 5.8%). No difference was observed for sperm DNA damage assessment using a 0.2 m (15.5 +/- 3.2%) or 0.3 m (14.9 +/- 3.3%) concentration of Na(2)HPO(4).7H(2)O in the AO staining solution. Frozen-thawed semen samples showed increased damage to sperm DNA under both Carnoy's (fresh: 10.9 +/- 1.3%; frozen: 30.8 +/- 2.9%; P < 0.05) and Diff Quik fixation (fresh: 6.2 +/- 0.8; frozen: 17.1 +/- 2.5%P < 0.05). Present data also showed that spermatozoa from some individuals are more prone to DNA damage after freezing and thawing procedures than others. In conclusion, Carnoy's fixative provides a better predictive value for DNA damage to sperm using AO staining. Additionally, cryopreservation increased damage to the sperm DNA.

Acetic Acid↗
PubMed2003

Sephadex and sephadex ion-exchange filtration improves the quality and freezability of low-grade buffalo semen ejaculates.

The effect of sephadex and sephadex ion-exchange filtration on the improvement in quality and freezability of low-grade buffalo semen ejaculates was assessed. Two types of filtration columns were used: one containing only sephadex G-10 (FS) and the other sephadex G-10 along with ion-exchangers (diethyl amino ethane-52 (DEAE-52) cellulose and carboxy methyl-52 (CM-52) cellulose; FS+IE). Unfiltered samples served as controls. Semen ejaculates extended in Tris-citric acid (1:4) (n=16; initial motility 40-50%) were filtered at the rate of 1.5 ml/min under negative pressure at room temperature (28-30 degrees C). The mean recovery rate (%) of motile spermatozoa in the FS (85.9+/-1.51) and FS+IE (77.10+/-2.28) filtrates did not differ significantly. Percentages of sperm motility, normal acrosomes, and intact plasma membranes were highest (P<0.05) in FS+IE, intermediate (P<0.05) in FS and lowest (P<0.05) in controls at the three stages of cryopreservation (postfiltration final dilution, after equilibration, and after freezing). Mean sperm abnormalities were lowest (P<0.05) in the filtrates of FS+IE, moderate (P<0.05) in FS and highest in controls at all stages of freezing. Compared to dilution and equilibration, freezing greatly reduced (P<0.05) the overall percent motility, normal acrosomes and intact plasma membranes. The spermatozoa eluted through FS+IE columns proved more resistant (P<0.05) in bearing dilution, equilibration, freezing and thawing stresses than the spermatozoa from FS and control samples. It is concluded that filtration systems containing an FS+IE column can effectively enhance the quality and freezability of extended, low quality buffalo semen.

Acrosome↗
PubMedDate not supplied

Mushroom cryopreservation and its effect on survival, yield and genetic stability.

Mycelial stock cultures of Agaricus bisporus, A. Bitorquis, Pleurotus flabellatus, P. Sajor-caju, P. Ostreatus, P. Sapidus, Auricularia polytricha, Lentinula edodes, Morchella esculenta and Volvariella volvacea were maintained by frequent subculturing at an interval of two months and separately as wheat grain spawn in liquid nitrogen with 15 percent glycerol. Preservation of mushroom stock cultures as wheat grain spawn under liquid nitrogen proved to be the better method of maintenance. The percent recoveries of stored samples were unchanged from the first recovery after six months to the last recovery after 42 months in nine out of 11 stock cultures preserved under liquid nitrogen. However, a marginal decline in survival of 10 % was recorded in Auricularia polytricha and Volvariella volvacea. Yields before preservation of mushroom stock cultures and after 30 months of preservation exhibited static biological efficiency and fruitbody weight. The comparison of Random Amplified Polymorphic DNA (RAPD) and Internal Transcribed Spacers (ITS) PCR amplified products did not exhibit DNA fragment variation in banding patterns at the intraspecific level during preservation of stock cultures by either method. The modified cryopreservation protocol and experimental demonstration of genetic stability of stock cultures reported here validate the use of mushroom cryopreservation techniques and supports studies on genetic stability of preserved biological materials.

Agaricales↗
PubMed1995

Lactate dehydrogenase-release assay: a reliable, nonradioactive technique for analysis of cytotoxic lymphocyte-mediated lytic activity against blasts from acute myelocytic leukemia.

Treatment of patients in remission of acute myelocytic leukemia using immunotherapy with interleukin 2 is a new approach to prolonging remission duration in this disease. As an important mechanism for the pathophysiology of eradication of residual myelocytic blast populations, activation of cytotoxic effector lymphocytes has frequently been discussed. However, the associated immunological research has been complicated to some extent, because in conventional chromium 51-release assays, blast cells frequently fail to incorporate sufficient amounts of 51Cr and/or spontaneously release high amounts of 51Cr. Recently, we established a culture system which promotes the outgrowth of cytotoxic T lymphocytes in bone marrow-derived mononuclear cells cultured in IL-2. To study cytotoxicity and the responsible mechanisms of the obtained T-cell lines and clones, we modified a previously described cytotoxicity assay, based on the release of lactate dehydrogenase (LDH-release assay) for use in cryopreserved blasts obtained from the bone marrow of patients with acute myelocytic leukemia. Using this assay, we were able to detect cytotoxicity of IL-2-activated peripheral blood lymphocytes from three healthy controls against a number of blast samples obtained from the bone marrow of patients with AML (up to more than 40% lysis at an effector target cell ratio of 20:1). However, a minority of AML blasts seem to be resistant to lysis by IL-2-activated lymphocytes. In bone marrow-derived T-cell lines from patients with AML we detected lytic activity against autologous blasts in three of seven cases tested by LDH release, ranging from 29 to 63% at an effector target ratio of 10:1. Additionally, T-cell clones with different phenotypes were established which were able to mediate cytotoxicity against autologous blast cells. Thus, cytotoxicity against freshly isolated blasts from patients with acute myelocytic leukemia can be analyzed reliably, reproducibly, and without the use of isotopes by the LDH-release assay.

Blast Crisis↗
PubMedDate not supplied

Strategies for "minimal growth maintenance" of cell cultures: a perspective on management for extended duration experimentation in the microgravity environment of a Space station.

How cells manage without gravity and how they change in the absence of gravity are basic questions that only prolonged life on a Space station will enable us to answer. We know from investigations carried out on various kinds of Space vehicles and stations that profound physiological effects can and often to occur. We need to know more of the basic biochemistry and biophysics both of cells and of whole organisms in conditions of reduced gravity. The unique environment of Space affords plant scientists an unusual opportunity to carry out experiments in microgravity, but some major challenges must be faced before this can be done with confidence. Various laboratory activities that are routine on Earth take on special significance and offer problems that need imaginative resolution before even a relatively simple experiment can be reliably executed on a Space station. For example, scientists might wish to investigate whether adaptive or other changes that have occurred in the environment of Space are retained after return to Earth-normal conditions. Investigators seeking to carry out experiments in the low-gravity environment of Space using cultured cells will need to solve the problem of keeping cultures quiescent for protracted periods before an experiment is initiated, after periodic sampling is carried out, and after the experiment is completed. This review gives an evaluation of a range of strategies that can enable one to manipulate cell physiology and curtail growth dramatically toward this end. These strategies include cryopreservation, chilling, reduced oxygen, gel entrapment strategies, osmotic adjustment, nutrient starvation, pH manipulation, and the use of mitotic inhibitors and growth-retarding chemicals. Cells not only need to be rendered quiescent for protracted periods but they also must be recoverable and further grown if it is so desired. Elaboration of satisfactory procedures for management of cells and tissues at "near zero or minimal growth" will have great value and practical consequences for experimentation on Earth as well as in Space. All of the parameters and conditions and procedural details needed to meet all the specific objectives will be the basis of the design and fabrication of cell culture units for use in the Space environment. It is expected that this will be an evolutionary process.

Bioreactors↗
PubMed1980

Immunological conditioning of bone marrow for autotransplantation in childhood acute lymphoblastic leukaemia.

Samples of bone marrow from 32 leukaemic children were removed during remission and stored in liquid nitrogen for retransplantation during relapse. Subsequently two children in advanced stages of common acute lymphoblastic leukaemia (cALL) were transplanted with their own cryopreserved marrow cells, after intensive combination chemotherapy and high doses of radiation therapy. Before grafting, the marrow cells were treated with purified heterologous antibodies prepared against cALL antigens, to remove any residual tumour cells. The antibodies showed high cytotoxic activity against leukaemic cells of cALL type without interfering with normal haemopoietic stem cells. Evidence of take was obtained in one patient, who died on day 7 with cardiac failure. In the other patient the dose of nucleated marrow cells grafted was 1.9 x 10(8)/kg (86 000 CFU-C/kg). The patient achieved complete haematological recovery on day 27 and a normal platelet count after day 50 and is now in complete remission. Marrow cells collected during remission and treated with antileukaemic antibodies can repopulate bone marrow after conditioning of the recipient with high doses of radiation.

Antibodies, Neoplasm↗
PubMed2006

Arsenic exposure is associated with decreased DNA repair in vitro and in individuals exposed to drinking water arsenic.

The mechanism(s) by which arsenic exposure contributes to human cancer risk is unknown ; however, several indirect cocarcinogenesis mechanisms have been proposed. Many studies support the role of As in altering one or more DNA repair processes. In the present study we used individual-level exposure data and biologic samples to investigate the effects of As exposure on nucleotide excision repair in two study populations, focusing on the excision repair cross-complement 1 (ERCC1) component. We measured drinking water, urinary, or toenail As levels and obtained cryopreserved lymphocytes of a subset of individuals enrolled in epidemiologic studies in New Hampshire (USA) and Sonora (Mexico). Additionally, in corroborative laboratory studies, we examined the effects of As on DNA repair in a cultured human cell model. Arsenic exposure was associated with decreased expression of ERCC1 in isolated lymphocytes at the mRNA and protein levels. In addition, lymphocytes from As-exposed individuals showed higher levels of DNA damage, as measured by a comet assay, both at baseline and after a 2-acetoxyacetylaminofluorene (2-AAAF) challenge. In support of the in vivo data, As exposure decreased ERCC1 mRNA expression and enhanced levels of DNA damage after a 2-AAAF challenge in cell culture. These data provide further evidence to support the ability of As to inhibit the DNA repair machinery, which is likely to enhance the genotoxicity and mutagenicity of other directly genotoxic compounds, as part of a cocarcinogenic mechanism of action.

Adult↗
PubMed1992

Anti-CD3 + interleukin-2 stimulation of marrow and blood: comparison of proliferation and cytotoxicity.

The proliferation and in vitro cytolytic activity of interleukin-2 (IL-2)-activated and anti-CD3 + IL-2-stimulated marrow mononuclear cells (MMC) and peripheral blood mononuclear cells (PBMC) were studied. Samples from 8 normal donors, 15 patients with acute lymphoblastic leukemia (ALL), and 7 patients with non-Hodgkin's lymphoma (NHL) in remission were cultured in IL-2 (100 U/mL) or IL-2 (100 U/mL) plus anti-CD3 (10 ng/mL). MMC as well as PBMC samples demonstrated significant synergy between IL-2 and anti-CD3 in the promotion of proliferation as measured by 3H thymidine incorporation on day 5 (P less than .001) or fold increase in cell number on day 14. Cryopreserved marrow specimens had equally rapid proliferation as fresh MMC when cultured in the presence of anti-CD3 + IL-2. Anti-CD3 concentrations of 3, 11, 33, and 100 ng/mL augmented proliferation similarly in the presence of IL-2 (0.1 to 100 U/mL). Mean fold increases in cell number of both marrow- and blood-derived cultures after 14 days were significantly higher for anti-CD3 + IL-2-stimulated cultures compared with cultures stimulated with IL-2 only (50- to 200-fold increase in cell number; P = .01). Comparison of remission MMC and PBMC from ALL and NHL patients with normal controls showed equivalent growth rates of activated cultures at 7, 14, and 21 days. Marrow purging with immunotoxin anti-CD19 pokeweed antiviral protein plus 4HC had no significant effect on proliferation of anti-CD3 + IL-2-stimulated MMC cultures in patients with ALL. Cytolytic activity of IL-2- and IL-2 + anti-CD3-activated PBMC and MMC cultures was assessed in 51Cr release assays using K562 (natural killer ([NK]-sensitive), Daudi (Burkitt's lymphoma-, NK-resistant), and Nalm-6 (ALL-, lymphokine-activated killer [LAK]-resistant) cell lines and cryopreserved ALL blasts. Cytolytic activity on a per-cell basis (percent cytotoxicity at an effector:target ratio of 30:1) was similar in IL-2-activated PBMC- and MMC-derived cultures from ALL patients. MMC activated with anti-CD3 plus IL-2 killed Daudi significantly less well than IL-2-activated cultures on days 12 and 19 (P = .03); no significant differences were observed in lysis of LAK-resistant Nalm-6 or cryopreserved ALL blast targets. Dose response of anti-CD3 augmentation of Daudi and Nalm-6 killing was different in IL-2- and IL-2 + anti-CD3-stimulated cultures.(ABSTRACT TRUNCATED AT 400 WORDS)

Bone Marrow↗
PubMed1998

Quantification of apoptosis by the Abbott CD4000 hematology analyzer.

We propose a simple and fast method of detecting apoptosis using an automated hematology analyzer. Detection is based on cellular optical light scatter properties and demonstration of the membrane fragility which characterizes cells undergoing the process of apoptosis. As part of it's routine leucocyte differential analysis, the Abbott Cell-Dyn 4000 collects multi-angle cellular light scatter data. In addition red fluorescence (FL3) emitted by cells following propidium iodide labeling is collected. This provides quantitation of both the erythroblast count and a leukocyte viability index (WVF). Fresh or cryopreserved peripheral blood cells from 17 B-chronic lymphocytic leukemia (B-CLL) patients were incubated in presence of theophylline, fludarabine or in medium alone. After 36-hrs of culture the percentage of apoptotic cells of the sample was determined from the parameters of the CD 4000 described above and thereafter this was compared with reference methods for estimation of apoptosis. The reference methods used were in situ detection of cell death on slides (TUNEL test) and also flow cytometry (Annexin V). Results showed an excellent correlation between the 3 techniques. This rapid, easy and reliable method of quantifying apoptosis may be very useful means of routinely predicting the response to chemotherapy.

Apoptosis↗
PubMed1990

"Two-step" freezing of individual blastomeres from mouse eight-cell embryos.

Eight-cell embryos recovered from superovulated C57BL/6J mice were disaggregated into individual 1/8 blastomeres. For cryopreservation of blastomeres a simple "two-step" technique based on direct transfer of blastomeres into the final concentration of DMSO (0.5 to 1.5 M) and exposure of plastic straws with blastomeres to -25 degrees C for 10 min before immersion into liquid nitrogen was used. The samples stored for 2 to 60 days were thawed at different temperature (20 to 80 degrees C) and intact 1/8 blastomeres were transferred into DMSO-free medium. The survival of blastomeres in individual groups varied from 52% to 90% according to the combination of DMSO concentration, exposure time to DMSO prior to freezing and temperature of thawing bath. Intact frozen-thawed 1/8 blastomeres were able to undergo cleavage and aggregates constructed from 4, 6, 7, and 8 blastomeres developed throughout preimplantation period. Of the aggregates constructed 1 to 2 h after thawing, 78% to 93% reached the blastocyst stage within 54 h of in vitro culture. Possible use of frozen-thawed 1/8 blastomeres in nuclear transplantation experiments and genetic manipulation on mammalian embryos is discussed.

Animals↗
PubMedDate not supplied

Penile vibratory stimulation and electroejaculation before anticancer therapy in two pubertal boys.

PURPOSE: Because more than 70% of children with cancer become long-term survivors, more emphasis is put on reducing late effects. Cryopreservation of semen and the intracytoplasmic sperm injection technique makes it possible to obtain pregnancy with very poor sperm quality. Two new semen retrieval methods are described that are applicable in pubertal boys with a fertility potential, although not psychologically ready to produce a semen sample, who are likely to become infertile because of anticancer therapy. PATIENTS AND METHODS: Two pubertal boys (aged 14 and 15 years) had a late testicular relapse of pre-B acute lymphoblastic leukemia and Hodgkin disease, stage II, respectively. In patient 1, penile vibratory stimulation (PVS) was tried under general anesthesia without success and electroejaculation (EEJ) was performed. Before alkylating chemotherapy and testicular irradiation, PVS was performed with success in patient 2. RESULTS: An antegrade ejaculate of 0.7 ml with 1% motile spermatozoa and an retrograde ejaculate with 1.6 x 10(6)/ml spermatozoa (5% with fair motility) was obtained from patient 1. An antegrade ejaculate of 1.5 ml with 2.5 x 10(6)/ml spermatozoa (29% with fair motility) was obtained from patient 2. CONCLUSIONS: PVS should be the first choice of treatment because it is noninvasive, simple, and easily applied. Because EEJ requires general anesthesia, it should be used as a second option.

Adolescent↗
PubMed2002

Follicle development in grafted mouse ovaries after cryopreservation and subcutaneous transplantation.

OBJECTIVE: Our aim was to determine the impact of freezing, thawing, and subcutaneous transplantation on follicular development in grafted mouse ovaries. STUDY DESIGN: The mice were divided into 3 groups: control (group 1), frozen-thawed grafting (group 2), and frozen-thawed grafting with human menopausal gonadotropin injection (group 3). After freezing and thawing, the ovaries were transplanted into the subcutaneous tissue. Two weeks after transplantation, grafted ovaries and blood samples were collected. RESULTS: Ovaries from group 3 contained significantly more follicles (246 +/- 43 follicles) than group 2(P <.05). The pattern and intensity of Cx37 immunohistochemical staining was similar in all groups. Follicle-stimulating hormone concentrations were significantly decreased in group 2 after ovarian grafting. CONCLUSION: In mice, gonadotropin treatment before subcutaneous grafting improved the survival of growing follicles. Subcutaneous ovarian transplantation may restore ovarian function and could obviate many of the problems that are related to ovarian banking for humans.

Animals↗
PubMedDate not supplied

Cryopreservation of plant germplasm using the encapsulation-dehydration technique: review and case study on sugarcane.

Encapsulation-dehydration is a cryopreservation technique based on the technology developed for producing synthetic seeds, i.e. the encapsulation of explants in calcium alginate beads. Encapsulated explants are then precultured in liquid medium with a high sucrose concentration and partially desiccated before freezing. Encapsulating the explants allows the subsequent application of very drastic treatments including preculture with high sucrose concentrations and desiccation to low moisture contents which would be highly damaging or lethal to non-encapsulated samples. An encapsulation-dehydration protocol comprises the following steps: pretreatment, encapsulation, preculture, desiccation, freezing and storage, thawing and regrowth. Encapsulation-dehydration has been applied to around 40 different plant species. The optimization of the successive steps of the encapsulation-dehydration protocol is illustrated for sugarcane apices.

Cryopreservation↗
PubMedDate not supplied

Penetration of human spermatozoa through the zona pellucida of nonviable human oocytes.

OBJECTIVE: The purpose of this study was to evaluate the penetrating capacity of human sperm through the intact zonae of nonviable human oocytes and the influence of oocyte storage conditions and gamete coincubation times on penetration rates. METHODS: Immature oocytes were obtained from surgically removed ovarian tissue and from in vitro fertilization (IVF) and subjected to four storage conditions: 1) storage at 4 degrees C for up to 48 hours in culture medium (refrigeration, n = 53), 2) cryopreservation in 1.5 mol/L propanediol with storage at -196 degrees C (PrOH; n = 49), 3) cryopreservation in 2.0 mol/L dimethylsulfoxide with storage at -70 degrees C (DMSO; n = 20), and 4) storage in a hypertonic salt solution at 4 degrees C (salt; n = 30). Semen was obtained from fertile donors (n = 7) and after a wash and swim-up, samples were adjusted to 500,000 motile sperm/mL. Zonae and sperm were coincubated in 100-microL drops under oil, and penetration was assessed at 5 and 20 hours. Penetration was evidenced by visualization of at least one sperm head in the perivitelline space at 200x magnification. RESULTS: Zona penetration rates were 50.9, 32.7, 23.3, and 10.0% for refrigeration, PrOH, salt, and DMSO storage, respectively (P < .004). There was a significantly higher number of sperm in the perivitelline space after 20 hours' coincubation (3.8 +/- 0.2) compared to 5 hours (0.8 +/- 0.1) (P < .0006). This represented an increase in the penetration rate from 31.2% (34 of 109) at 5 hours to 37.6% (41 of 109) at 20 hours, although the difference was not significant. CONCLUSIONS: These results demonstrate that the zona pellucida of nonviable human oocytes can be used in a zona penetration assay and that refrigerated and PrOH-frozen intact zonae have the highest rates of penetration. The decrease in penetrability seen in DMSO and salt-stored oocytes may be due to alterations occurring in the zona pellucida.

Cell Survival↗
PubMed1996

Digital photogrammetric quantification of surface area and volume on scanning electron micrographs of frozen hydrated lung tissue.

A digital video plotter (DVP, Leica), the personal computer equivalent of an analytical plotter, was used to measure the coordinates of points chosen from stereo pair images of the surface of frozen hydrated lung imaged at magnifications of 2000 and 5000 X with a low-temperature scanning electron microscope (SEM). Rat lung tissue was frozen in vivo with a liquid nitrogen cryoprobe under carefully controlled physiologic conditions. At slow freezing rates, water in the aqueous layer at the surface of the lung segregates into ice crystals (dendrites) which branch in the plane of the surface. Coordinates of points on dendrite surfaces were measured by the DVP and passed to TERRAMODEL (Plus III Software, a land modeling program) where they were used to generate a three-dimensional model, from which surface area and planimetric area of the lung surface were calculated. Additional measurements were made at the top and bottom of the ice structures and a Basic language program was written to calculate the volume of ice on the lung surface. Digital photogrammetry coupled with low-temperature SEM of frozen samples allows measurement of water and water-containing microstructures ubiquitous in biology.

Animals↗
PubMed1999

Freeze-drying of red blood cells at ultra-Low temperatures.

The hemolysis of human red blood cells (RBCs) after freeze-drying and resuspension depends on the vacuum-drying temperature. In an experimental study, RBCs were first solidified based on a modified high-yield cryopreservation protocol in the presence of hydroxyethyl starch and maltose. Afterward, they were vacuum-dried in a special low-temperature freeze-drying device at selected shelf temperatures between -5 and -65 degrees C. Subsequently, the dried samples were resuspended in an isotonic, phosphate-buffered saline solution. The hemolysis was determined according to a modified saline stability test. It decreases with a decreasing shelf temperature until a minimum is reached at -35 degrees C. A further decrease of the shelf temperature has no beneficial effect; the hemolysis even increases. To interpret these results, we assume that the hemolysis depends on two contrary damaging effects: (1) the higher the shelf temperature, the higher the probability of structural damages occurring during drying; (2) the lower the shelf temperature, the lower the driving force for water transport; this may lead to an incomplete intracellular dehydration which means that the cells are not in a glassy state at ambient temperature.

Blood Preservation↗
PubMed2003

[Histological and cytopathological cancer specimens: good practice in operating room].

Concerning good practice for the management of histological cancer specimen, the main recommendations concern: (1) the quality of transmission in information between professionals; (2) the necessity of fresh, unfractionated, oriented surgical samples; (3) the importance of an appropriate storage condition for samples collected for extemporaneous examination, freezing or cell culture; (4) the quality of the deep freezer at temperatures of -80 degrees C or liquid nitrogen for frozen samples storage; (5) the importance of fixing tissues shortly after sample collection in buffered Formal solution in order to prevent cell lysis.

Benchmarking↗
PubMed2006

Deciding the fate of supernumerary frozen embryos: a survey of couples' decisions and the factors influencing their choice.

OBJECTIVE: To investigate the decisions that couples make regarding supernumerary frozen embryos, the factors influencing these decisions, and the degree of difficulty involved in reaching a decision; and to canvass attitudes toward donating embryos to stem-cell research. DESIGN: Anonymous postal survey. SETTING: A large, private IVF clinic in a major city in Victoria, Australia. PATIENT(S): A consecutive cohort of couples who contacted the Monash IVF clinic in relation to embryos in long-term storage. INTERVENTION(S): Subjects completed a survey regarding decisions about surplus frozen embryos. MAIN OUTCOME MEASURE(S): Couples' decision regarding supernumerary embryos and reasons for the decision, experience of deciding, and attitudes about embryo donation for stem-cell research. RESULT(S): Forty percent (123/311) returned completed questionnaires. The most common decision was donation to research (42%). Altruistic motives and desire not to waste embryos were determinants of embryo donation. Determinants of disposal were not wanting a full sibling to existing children and opposition of embryo research. Forty-five percent found deciding distressing. The majority (69%) approved of embryo donation to stem-cell research. CONCLUSION(S): Most couples preferred embryos to come to some use rather than being disposed of. Almost half the sample reported finding the decision making distressing. A majority approved of embryo donation for stem-cell research.

Adult↗