Background: Sperm cryopreservation becomes a relatively routine process in assisted reproductive centers. However, there must be ensured quality of washed human normal sperm and cryopreservation to successful fertilization. Objective: To evaluate the quality of washed human normal sperm after cryopreservation. Subjects and method: 30 normal semen samples, each sample was divided into two parts for washed and unwashed spermatozoa. All samples were cryopreserved in 1, 2 and 30 days. Evaluating and comparing the quality of sperm before and after which washed, pre-cryopreservation and post-cryopreservation between the groups were performed. Results: The quality of sperm after washing was more significantly improved than before washing. Post-cryopreservation, the quality of sperm was reduced time by time but within an accepted limitation. There was not a significant difference between the two ways of preparation before cryopreservation. Conclusions: The quality of sperm at post-cryopreservation was reduced (both washed sperm and unwashed sperm). The quality of washed sperm is reduced continuously with time, but there was no difference between the two studied groups.
Washed sperm ; sperm cryopreservation

Introduction: Successful cryopreservation of spermatozoa must ensure normal newborns after the preservation time. This method frequently can potentially contain cross-infected risks during the cryopreservation process in the liquid nitrogen environment (such as HCV, HIV). A number of researchers reveal that these risks can be eliminated by washing spermatozoa before cryopreservation. However, the problem is whether cryopreservation of washed spermatozoa still retains its morphology and function or not? \r\n', u'Objectives: To evaluate the change of sperm morphology characteristics after which washed sperm cryopreserved from normozoospermia. \r\n', u'Subjects and method: 30 normal semen samples; each sample was divided into two aliquots of washed and unwashed spermatozoa. All samples were cryopreserved in stages of 1, 2 and 30 days. We compared the percentage of spermatozoa with normal morphology before and after which was washed, pre - cryopreservation and post - cryopreservation between the groups. \r\n', u'Results: The percentage 0 spermatozoon with normal morphology after washing was more significantly increased than prior to washing. Post - cryopreservation, this percentage was reduced time by time but acceptable. There is no significant difference between the two ways of preparation before cryopreservation. The percentage of spermatozoa with abnormal head and neck increased significantly after cryopreservation. \r\n', u'Conclusion: The percentage of spermatozoa with normal morphology post - cryopreservation was reduced in both washed sperm and unwashed sperm samples. This percentage was reduced time by time, but there is no difference between the two groups studied. \r\n', u'\r\n', u'
Sperm washing ; Sperm cryopreservation ; Sperm morphology

PURPOSE: Intrinsic milieu of seminal plasma is important in maintaining the survival and fertilizing capacity of normal sperm. Nevertheless, there are still debates concerning the addition of seminal plasma during sperm preparation and cryopreservation. The purpose of this study is to investigate the effects of autogenous seminal plasma on the recovery of frozen fertile and infertile human sperm. MATERIALS AND METHODS: Semen from 10 fertile volunteers and 10 infertile patients was used. Sperm pellet from each group was resuspended and cryopreserved with 0.5ml cryoprotectant. Cryoprotectant was added with 100% (group I), 75% (group II), 50% (group III), 25% (group IV) and 0% (group V) seminal plasma according to each group. After 3 days, each vial was thawed and examined about sperm concentration, motility and morphology by CASA and Makler chamber. Sperm vitality was examined by eosin-nigrosin stain. RESULTS: The recovery rates of sperm motility and vitality after thawing in normal volunteers were higher in group I and II than other groups with statistical significance (p<0.05). However there was no difference between groups in sperm concentration and morphology (p>0.05). In the infertile patients all factors had no difference between groups. CONCLUSIONS: We concluded that the sperm quality following the addition of autogenous seminal plasma in cryoprotectant was improved in normal fertile men. This results may give a theoretical basis to support the clinical use and application for the seminal plasma to enhance the sperm quailty.
Cryopreservation* ; Healthy Volunteers ; Humans* ; Male ; Semen* ; Sperm Motility ; Spermatozoa* ; Volunteers

OBJECTIVETo observe sperm motion parameters in pre-freeze and post-thaw semen samples using computer-assisted sperm analysis (CASA) system.

METHODSSemen analyses of 238 samples before freezing and after thawing were separately performed by Hamilton-Thorne Sperm Analyzer.

RESULTSSperm motility in post-thaw samples was significantly decreased. There was significant correlation and difference between pre-freeze and post-thaw samples in sperm motion parameters, including average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), straightness (STR) and linearity (LIN), except heat cross frequency (BCF). The percentage of sperm movement velocity parameters (VAP, VSL and VCL) and moving pattern parameters (ALH) significantly decreased, while that of LIN and STR significantly increased in post-thaw samples.

CONCLUSIONCASA system is of clinically applied value and is a useful tool for evaluating sperm motion parameters in pre-freeze and post-thaw semen samples.

Cryopreservation ; Diagnosis, Computer-Assisted ; Freezing ; Humans ; Male ; Sperm Motility

Sperm cryopreservation has revolutionized the field of assisted reproduction. However, it causes cryodamage to the structure and function of spermatozoa during the freezing-thawing process. Optimization of sperm cryopreservation is necessary for the preservation of male fertility. Cryodamage can be reduced effectively by such methods as improvement of semen quality before freezing, spermatozoal selection, addition of optimal cryoprotectants and application of appropriate thawing techniques. Recent studies focus on cryobiology, improvement of freezing-thawing methods, especially for poor quality semen, and evaluation criteria for post-thaw spermatozoa.
Cryopreservation ; methods ; Humans ; Male ; Semen Preservation ; methods ; Sperm Count ; Sperm Motility ; Spermatozoa ; cytology ; physiology

OBJECTIVETo investigate sperm function indexes that can be used to effectively evaluate the sperm donors' fertility so as to select healthy post-thaw semen samples and improve the success rate of assisted reproductive technology.

METHODSAccording to the pregnancy outcomes, we divided 40 donor semen samples into a high-fertility group (n = 20) and a low-fertility group (n = 20). We measured and compared the concentration, progressive motility, morphology, acrosome intactness, DNA integrity and mitochondrial membrane potential (MMP) of the post-thaw sperm between the two groups.

RESULTSThere were statistically significant differences between the high- and low-fertility groups in the percentages of morphologically normal sperm ([18.50 +/- 6.10]% vs [14.42 +/- 6.44]%, P < 0.01), acrosome intactness ([86.17 +/- 4.49]% vs [80.04 +/- 7.52]%, P < 0.05) and DNA fragmentation index ([9.21 +/- 3.22]% vs [15.72 +/- 8.20]%, P < 0.05), but not in MMP ([56.75 +/- 18.80]% vs [52.23 +/- 18.86]%, P > 0.05). A significantly positive correlation was found between MMP and sperm motility (r = 0.760, P < 0.05), but not between other sperm functions and sperm concentration and motility.

CONCLUSIONSperm concentration, motility, morphology, acrosome intactness rate and DNA integrity contribute effectively to the evaluation of the fertilization capacity of post-thaw donor semen samples.

Adult ; Cryopreservation ; Female ; Fertilization ; Humans ; Male ; Pregnancy ; Semen Preservation ; Sperm Banks ; Sperm Count ; Sperm Motility ; Spermatozoa ; physiology

OBJECTIVETo investigate the effect of pre-freezing equilibration on the cryo-survival of human sperm and to optimize the protocol of direct fumigation for the freeze-thawing of human sperm.

METHODSWe collected 50 semen samples from healthy donors, each subjected to cryopreservation with 3 different methods: non-equilibration freezing (Group A), 10-min equilibration at room temperature before freezing (Group B), and 10-min equilibration at 4 degrees C before freezing (Group C). We examined all the post-thaw semen samples by computer-assisted semen analysis for the sperm motility parameters, and detected the sperm vitality and deformity index (SDI).

RESULTSThe recovery rate of progressive sperm motility was (61.88 +/- 16.94)% in Group C, remarkably higher than in A ([48.61 +/- 16.44]%) and B ([49.41 +/- 13.77]%) (P < 0.05), but with no significant difference between the latter two. And there were no significant differences in sperm vitality and SDI among the three groups.

CONCLUSIONTen-minute equilibration at 4 degrees C before freezing can evidently improve the progressive motility of sperm in addition to its advantages of easy operation and controllable experimental condition.

Adult ; Cryopreservation ; methods ; Humans ; Male ; Semen Analysis ; Semen Preservation ; methods ; Sperm Banks ; Sperm Count ; Sperm Motility ; Young Adult

OBJECTIVETo investigate the relationship of seasonal variation with pre- and post-thaw semen parameters as well as the cryosurgical of human spermatozoa.

METHODSA total of 6 414 semen samples were collected from 1 135 donors aged 22 - 32 years by Zhejiang Human Sperm Bank, and divided into spring, summer, autumn and winter groups according to the time of collection. All the samples underwent routine seminal analysis, and the sperm parameters were compared between different seasons. The sperm specimens were cryopreserved in aliquots and analyzed after thawing.

RESULTSThe semen volume was (2.92 +/- 1.17) ml in spring, significantly higher than in summer, autumn and winter ([2.71 +/- 1.07 ], [2.74 +/- 1.15] and [2.83 +/- 1.15] ml, P < 0.05). Sperm density was the highest in autumn ([105.60 +/- 39.76] x 10(6)/ml) as compared with the other three seasons ([101.18 +/- 40.16] x 10(6)/ml, [93.54 +/- 35.10] x 10(6)/ml, and [101.29 +/- 38.37] x 10(6)/ml, P < 0.05). The sperm progressive motility was the highest in spring ([58.49 +/- 10.04] %) and the cryosurgical of sperm the lowest in summer, with statistically significant differences from the other groups (P < 0.05).

CONCLUSIONSeasonal variations affect human semen quality and cryosurgical of sperm. The semen volume, the percentage of progressive motile sperm, the cryosurgical of sperm, and the post-thaw density of progressive motile sperm are higher in spring than during the rest of the year.

Adult ; Cryopreservation ; Humans ; Male ; Seasons ; Semen Analysis ; Semen Preservation ; Sperm Banks ; Sperm Count ; Sperm Motility ; Tissue Donors ; Young Adult

There are not much reports concerning with clinical results using frozen-thawed testivular sperm in ICSI program. It is speculated that the necessity of cryopreservation of testicular sperm to avoid repeating surgical procedure for obtaining sperm for ICSI. This study was carried out to confirm whether frozen-thawed testicular sperm could be fertilized and pregnancy could be achieved using embryos fertilized with frozen-thawed testicular sperm in ICSI program or not. Testicular sperm obtained from obstructive- or non-obstructive azoospermia patients were co-cultured for 3 days with Vero cells to improve sperm motility. By co-culturing with Vero cells for 3 days, O-ll% of sperm motility after thawing increased up to 8-42% after co-culturing. ICSI was performed using frozen-thawed, and co-cultured sperm with 66 oocytes obtained from 8 patients and 62 oocytes were survived and 49(79.0%) oocytes were fertilized normally. Embryo transfer was possible in 7 out of 8 patients, and pregnancy was achieved in 6 patients(85.7%). These results indicated that not only fresh testicular sperm but frozen-thawed testicular sperm can be used in ICSI program.
Azoospermia ; Cryopreservation ; Embryo Transfer ; Embryonic Structures ; Humans ; Oocytes ; Pregnancy* ; Sperm Injections, Intracytoplasmic* ; Sperm Motility ; Spermatozoa* ; Vero Cells

OBJECTIVETo evaluate the impact of sperm midpiece morphology observed under high-power microscope on embryo development following intracytoplasmic morphologically selected sperm injection.

METHODSMorphologically normal sperms from 57 patients undergoing intracytoplasmic sperm injection (ICSI) for male-factor infertility were selected microscopically (magnification of ×200 or 400) and subjected to motile sperm organellar morphology examination (MSOME) at high magnification of ×6000. According to the morphology of sperm medpiece, the sperms were divided into 3 groups, namely group A with a/b of 1-1.2, group B with a/b≥1.5, and group C with irregular morphology. The sperms in the 3 groups were intracytoplasmically injected in oocytes and the outcomes of the embryos were compared.

RESULTSGroups A, B, and C showed significant differences in the rate of ET-D3 top quality embryo (79.7% vs 55.6 % vs 33.3%) and implantation rate (43.2% vs 11.1% vs 0%), but not in the fertilization rate (73.3% vs 80.4% vs 63.5%), blastocyst formation rate (23.2% vs 22.2% vs 9.09%), cryopreservation rate (29.2% vs 25.0 % vs 13.0%), or D3 top quality embryo rate (35.3% vs 37.8% vs 18.8%).

CONCLUSIONSIn ICSI cycle, selecting morphologically normal sperms for intracytoplasic injection can increase the normal fertilization rate and top quality embryo rate on the transfer day and improve the implantation rate of the embryo.

Cryopreservation ; Embryo Implantation ; Embryonic Development ; Female ; Fertilization ; Humans ; Infertility, Male ; Male ; Oocytes ; Semen Analysis ; Sperm Injections, Intracytoplasmic ; Sperm Midpiece ; physiology