Genetic and epigenetic risks of intracytoplasmic sperm injection method

Pregnancies achieved by assisted reproduction technologies, particularly by intracytoplasmic sperm injection （ICSI） procedures, are susceptible to genetic risks inherent to the male population treated with ICSI and additional risks inherent to this innovative procedure. The documented, as well as the theoretical, risks are discussed in the present review study. These risks mainly represent thatconsequences of the genetic abnormalities underlying male subfertility （or infertility） and might become stimulators for the development of novel approaches and applications in the treatment of infertility. In addition, risks with a polygenic background appearing at birth as congenital anomalies and other theoretical or stochastic risks are discussed. Recent data suggest that assisted reproductive technology might also affect epigenetic characteristics of the male gamete, the female gamete, or might have an impact on early embryogenesis. It might be also associated with an increased risk for genomic imprinting abnormalities.

Clinical pregnancies and livebirths achieved by intracytoplasmic injection of round headed acrosomeless spermatozoa with and without oocyte activation in familial globozoospermia： case report

We report the successful outcome of intracytoplasmic sperm injection （ICSI） treatment in two siblings with familial globozoospermia. After controlled ovarian hyperstimulation and oocyte pick-up, retrieved oocytes were mechanically activated before ICSI and a fertilization rate of 33.3% was achieved in the first case. The second couple underwent ICSI without oocyte activation and a 9.1% fertilization rate was obtained. The transfer of two grade I embryos in the first couple and one grade I embryo in the second couple resulted in clinical pregnancies with healthy livebirths. It was concluded that the main problem of cases with globozoospermia is a low fertilization rate, and even though ICSI and oocyte activation can increase this rate it is not necessarily needed to achieve a pregnancy. （Asian J Androl 2008 Mar; 10： 332-336）

Pregnancies established through intracytoplasmic sperm injection (ICSI) using spermatozoa with dysplasia of fibrous sheath

Aim:Dysplasia of the fibrous sheath(DFS)is an anomaly found in asthenozoospermic patients with extremely lowor absent motility.In order to determine the efficacy of ICSI in these patients,a retrospective analysis of ICSI results inDFS patients has been done.Methods:Ten ICSI attempts were performed in 6 patients with diagnosis of Dysplasiaof the Fibrous Sheath studied by transmission and scanning electron microscopy.Results:In the cases studied,sperm concentration was(29.62±18.05)×10^(6)/mL,total motility was 1.14±1.31%.Progressive motility was 0%except for one case with 0.1%.One hundred and three preovulatory oocytes were obtained and 94 metaphaseⅡoocyteswere injected.Sixty-nine of them showed two pronuclei(fertilization rate:73.4%).Forty-nine embryos were ob-tained and 34 were transferred(mean:3.4 embryos per transfer).Five pregnancies were diagnosed byβ-hCG plasmalevel determinations that resulted to be one preclinical abortion,one clinical abortion and three deliveries.Anotherpregnancy(ongoing)was achieved from a cryopreserved embryo transfer.Conclusion:These results showed thatICSI provides a suitable solution for patients suffering from irreversible sperm defects such as DFS.Nevertheless,it ismandatory to inform couples of possible transmission risks to offspring,which are unknown at present.Only when theetiology of this problem is disclosed,it will be possible to assess the real genetic risk.

Intracytoplasmic sperm injection in cases with a history of in vitro fertilization failure

<abstract>Aim: To evaluate the effect of intracytoplasmic sperm injection (ICSI) in the management of cases with a history of conventional in vitro fertilization (IVF) failure. Methods: Two groups of patients, 19 with normal semen parameters and a history of IVF failure (metaphase Ⅱ oocytes: 0~30 %) and 28 with severe male factor infertility received ICSI technology during the same period. Ovarian stimulation was achieved by conventional procedure. Transvaginal ultrasound-guided oocyte collection was done 35~37 h after human chorionic gonadotrophin (hCG) injection. Only metaphase Ⅱ oocytes were selected for microinjection. Results: Fertilization was achieved with ICSI in all the patients. The fertilization rate (75.6 %±21.1 % vs. 73.9 %±19.2 %), cleavage rate (85.1 %±19.3 % vs. 82.7 %±22.1 %), clinical pregnancy rate per embryo transfer cycle (31.6 % vs. 28.6 %) and implantation rate per embryo (15.3 % vs. 14.4 %) did not differ significantly between the two groups. Conclusion: ICSI is a valuable method for couples with a history of IVF failure. These patients may have a similar ICSI result as in severe male infertility.

Fertilization of in vitro matured human oocytes by intracytoplasmic sperm injection (ICSI) using ejaculated and testicular spermatozoa

Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.

A nomogram predicting clinical pregnancy in the first fresh embryo transfer for women undergoing in vitro fertilization and intracytoplasmic sperm injection(IVF/ICSI) treatments

The extent to which factors affect the probability of clinical pregnancy in the first fresh embryo transfer after assisted conception is unknown.In order to examine the predictors of clinical pregnancy,a retrospective cohort study was launched between January 1,2013 and December 31,2016 in four infertility clinics including 19837 in vitro fertilization and intracytoplasmic sperm injection(IVF/ICSI)fresh cycles with known outcomes and relevant records.A multivariable logistic regression was used to select the most significant predictors in the final nomogram for predicting clinical pregnancy.Furthermore,the model was validated by an independent validation set and the performance of the model was evaluated by the receiver operating characteristic(ROC)curves along with the area under the ROC curve(AUC)and calibration plots.In a training set including 17854 participants,we identified that female age,tubal factor,number of embryos transferred,endometrial thickness and number of good-quality embryos were independent predictors for clinical pregnancy.We developed a nomogram using these five factors and the predictive ability was 0.66 for AUC(95%CI=0.64−0.68),which was independently validated in the validation set(AUC=0.66,95%CI=0.65−0.68).Our results show that some specific factors can be used to provide infertile couples with an accurate assessment of clinical pregnancy following assisted conception and facilitate to guide couples and clinicians.

Cumulative live birth rates of in vitro fertilization/intracytoplasmic sperm injection after multiple complete cycles in China

There were few studies of cumulative live birth rates(CLBRs) based on multicenter reproductive clinical data from the general Chinese population.Here we report a retrospective cohort study,including 14 311 women with17 315 cycles,in three reproductive centers to evaluate two estimated parameters of CLBRs with multiple transfer cycles of in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI) in a Chinese population.We found that CLBRs were related to female age and endometrial thickness.By the fourth transfer cycle,the conservative and optimal estimates of CLBRs were 52.95% and 77.30% in women under 30 years of age,and 18.17% and26.51% in those 37 years of age or older,respectively.The two estimates were 44.70% and 63.15% in women with endometrial thickness more than 7 mm,and 32.05% and 46.18% in those with less than 7 mm,respectively.In addition,body mass index(BMI),duration of infertility,and infertility diagnoses may also be related to CLBRs on certain conditions.The findings from this study on CLBRs after multiple transfer cycles of IVF/ICSI treatment on different conditions in the Chinese population should be beneficial to both infertile couples and clinicians.

Birth after intracytoplasmic sperm injection of ejaculated spermatozoa from a man with mosaic Klinefelter's syndrome

Aim:To report a birth after intracytoplasmic sperm injection (ICSI) of ejaculated spermatozoa from a man with mosaic Klinefelter's syndrome detected by fluorescence in situ hybridization (FISH) analysis.Methods:A 35-year- old man with a normal appearance consulted our hospital because of sterility over a 5-year period.Chromosome analysis showed low-incidence mosaic Klinefelter's syndrome.Using FISH,96 % hyperploidy of the lymphocytes was found.We examined the sex chromosome of the ejaculated spermatozoa.Using FISH,we examined 200 ejacu- lated spermatozoa and no hyperploidy was found.Results:The 33-year-old female partner of the male patient underwent an uncomplicated controlled ovarian hyperstimulation sequence using a combined recombinant-follicle stimulating hormone (rec-FSH) + human menopausal gonadotrophin (hMG) protocol,following late luteal phase pituitary down regulation.This culminated in the retrieval of seven oocytes,six of which were fertilized with ICSI. One ICSI attempt led to clinical pregnancy with a healthy baby girl.Conclusion:We report a male patient with low- incidence mosaic Klinefelter's syndrome whose ejaculated spermatozoa were identified as being haploid by FISH before ICSI,leading to the successful pregnancy of his wife and the birth of a healthy baby girl.

Intracytoplasmic sperm injection in the treatment of male infertility due to obstructive or non-obstructive azoospermia

Objective: To evaluate the effects of intracytoplasmic sperm injection (ICSI) ontreatment of infertility due to obstructive and non-obstructive azoospermia..Methods: A retrospective analysis of fertilization, cleavage, embryo implantationand pregnancy rates was done in 158 ICSI cycles including 112 obstructive azoospermiaand 46 non-obstructive azoospermia. Ovarian hyperstimulation and ICSI procedureswere performed by conventional protocol. The sperm was collected by percutaneous epi-didymal sperm aspiration (PESA) or testicular sperm extraction (TESE).Results:The fertilization rate (73.1% vs. 67.0%), cleavage rate (88.6% vs. 86.3%), embryo implantation rate (20.7% vs. 11.4%), clinical pregnancy rate per trans-fer cycle (35.7% vs. 19.6%) were obtained for obstructive and non-obstructiveazoospermia, respectively.Conclusion: The results revealed that in the cases of obstructive azoospermia, ferti-lization rate, embryo implantation rate and clinical pregnancy rate were significantlyhigher than those of non-obstructive azoospermia. But there was no significant differ-ence of the cleavage rate between two groups.

Effect of Vitamin D Supplementation on Intracytoplasmic Sperm Injection Outcomes: A Randomized Double-Blinded Placebo-Controlled Trial

OBJECTIVE: To assess the effect of vitamin D supplementation on enhancement of embryos implantation rates in intracytoplasmic sperm injection procedures. STUDY DESIGN: A randomized double-blinded controlled trial. SETTING: Assisted Reproduction Unit of Obstetrics and Gynecology Department, Faculty of Medicine, South Valley University, Egypt. DURATION: From December 1st 2017 to the end of October 2018. SUBJECTS and METHDS: Eighty infertile couples that underwent ICSI procedure were entrapped in this study. Patients were divided into two groups;Group I (Vitamin D group) = 40 patients (received vitamin D supplementation in the form of Vidrop oral drops 600 IU/day starting after ovum pickup) and Group II (placebo group) = 40 patients (received a placebo “normal saline oral drop”). RESULTS: There was a significantly increased embryo implantation rates (53.2% in vitamin D group versus 46.7% in placebo group), there was a mild statistically significant difference in the clinical pregnancy rates (45.9% in vitamin D group versus 39.4% in placebo group) with p-value < 0.05 and there was an improvement in ongoing pregnancy rate as there was a mild statistically significant difference in first trimester miscarriage rates (12.5% in vitamin D group versus 17.5% in placebo group) with p value CONCLUSIONS: Vitamin D supplementation could improve embryos implantation rates and rate of ongoing pregnancy by decline in the first trimester miscarriage rate.

Role of Dehydroepiandrosterone Supplementation in Improving Intracytoplasmic Sperm Injection Outcome for Women with Expected Poor Ovarian Response

Background: As regard to adjuvant supplementations, nowadays dehydroepiandrosterone (DHEA) is widely used all over the world and is considered to be a potential agent to ameliorate the assisted reproduction technologies outcomes of infertile women with poor ovarian reserve. Objective: To find out the role of DHEA supplementation in improving intracytoplasmic sperm injection (ICSI) outcome for infertile women with expected poor ovarian response in controlled ovarian stimulation. Setting: Assisted reproduction unit of Obstetrics and Gynecology Department, Faculty of Medicine, South Valley University, Egypt. Duration: From April 2016 to May 2018. Study Design: A randomized double-blinded controlled trial. Methods: One hundred and forty infertile women with expected poor ovarian response prepared for ICSI procedure were included in this study. Patients were divided into two groups;group I (DHEA group) included 70 patients received 25 mg DHEA 12 weeks prior to ICSI cycle and group II (placebo group) included 70 patients received a placebo. Results: There was a highly statistically significant difference in basal AFC at start of ICSI cycle in group I (who received DHEA supplementation for 12 weeks prior to ICSI procedure) than in group II (13.8 ± 5.3 versus 10.7 ± 4.6 respectively) with P < 0.001. There were mildly statistically significant differences between group I and group II as regard to increase in the number and quality of retrieved oocytes, increased in endometrial thickness, fertilization rate and embryo quality with p value < 0.05 but there was no statistically significant difference between the 2 groups as regard to pregnancy (chemical and clinical) rates (p value > 0.05). Conclusions & Recommendations: DHEA supplementations improved basal AFC, increased the number & quality of oocytes and increased quality of embryos in infertile patients with expected poor ovarian response in ICSI procedure. So DHEA supplementations could be an important adjuvant for infertile women with expected poor ovarian response in ICSI procedure.

In vitro maturation of human oocytes maintaining good development potential for rescue intracytoplasmic sperm injection with fresh sperm

BACKGROUND The outcomes of the use of commercial in vitro maturation(IVM)medium to culture immature oocytes obtained from conventional ovulation induction,followed by rescue intracytoplasmic sperm injection(RICSI),are not ideal.It is thus difficult to widely adopt this approach in clinical practice.Therefore,it is necessary to explore methods for improving the clinical outcome of IVM.AIM To study the effect of sperm on the developmental potential of in vitro-matured oocytes in conventional culture.METHODS This was a retrospective study of patients whose immature oocytes were harvested from conventional oocyte stimulation cycles and underwent ICSI at our hospital between June 2018 and August 2020.RICSI was performed using sperm collected on the day of oocyte harvest(old)and sperm collected on the day of RICSI(fresh)and oocytes matured in vitro after 24 h of culture in conventional medium.The rates of in vitro oocyte maturation,normal fertilization,normal cleavage,day-3 top-quality embryos,and useful blastocyst formation were compared between the two groups.RESULTS In total,102 germinal vesicle(GV)-stage immature oocytes were cultured in the old sperm group.In the fresh sperm group,122 GV-stage immature oocytes were collected and cultured in vitro for 24 h.There were no significant differences in the general conditions of males and females between the two groups(P>0.05).The oocyte maturation,normal fertilization,and normal cleavage rates of the old and fresh groups were 51.0%vs 55.7%,61.5%vs 64.7%,and 93.8%vs 93.2%,respectively.None of the rates differed significantly(P>0.05)between the two groups.However,the day-3 top-quality embryo and useful blastocyst rates of the old and fresh sperm groups were 16.6%vs 63.4%;6.67%vs 34.6%,respectively.The day-3 top-quality embryos and useful blastocyst rates of the old sperm group were significantly lower than those of the fresh group(P<0.05).CONCLUSION In vitro maturation with conventional culture medium combined with the use of fresh sperm collected on the day of RICSI is an easy-to-implement strategy for patients whose oocytes are completely or mostly immature.

Effect of Fasting of Ramadan on Infertile Women Undergoing In-Vitro Fertilization/Intracytoplasmic Sperm Injection Cycles: A Prospective Cohort Study

Objective: To determine the effects of fasting of Ramadan in patients undergoing in-vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles. Design: A prospective cohort study conducted during Ramadan. Setting: Ain Shams University IVF centre. Population: Three hundred fasting, infertile women undergoing their first trial of IVF/ICSI and 300 non-fasting matched controls undergoing the same procedure during Ramadan. Methods: Anxiety and depression were measured by the Hospital Anxiety and Depression Scale, at the start of the induction protocol. All the successful pregnant women were followed up until delivery. Main Outcome Measures: The primary outcome measure was the live birth rate. Results: The live birth rate was higher in the fasting group than in the non-fasting group;this was not significant (43% vs. 40%, P = 0.46). The fasting group needed higher doses and durations of induction. Embryo quality did not differ between both groups. The pregnancy complication rate among successful cases was higher in the fasting group (52.9% vs. 40.4%, P = 0.03);however, the frequencies of anxiety and depression were significantly lower than those in the non-fasting group (18% vs. 38%, P P < 0.00001, respectively). Conclusion: Fasting during Ramadan does not seem to significantly affect the IVF/ICSI outcome;however, it significantly decreased the anxiety and depression usually associated with these procedures.

体外受精/卵胞质内单精子注射反复种植失败的药物治疗研究进展

体外受精/卵胞质内单精子注射(in vitro fertilization/intracytoplasmic sperm injection,IVF/ICSI)反复种植失败(repeated implantation failure,RIF)是生殖医学领域的难题。目前,RIF药物治疗的研究较多,免疫因素在胚胎种植及妊娠维持中发挥重要作用,经验性免疫相关药物治疗可改善部分RIF患者妊娠结局,治疗药物包括:低分子量肝素(LMWH)、粒细胞集落刺激因子(G-CSF)、硫酸羟氯喹(HCQ)、环孢素A(CsA)、肿瘤坏死因子α(TNF-α)抑制剂、富血小板血浆(PRP)、外周血单个核细胞(PBMC)、静脉注射免疫球蛋白(IVIG)、脂肪乳剂等。对上述药物治疗的研究进展进行综述,以期为临床提供参考。

辅助生殖技术中精子DNA损伤的研究现状

精子DNA完整性对受精卵的形成至关重要,精子DNA损伤可能影响辅助生殖技术(assisted reproductive technology,ART)的成功率。精子DNA损伤主要包括单链和双链损伤,这些损伤可能在受精、胚胎质量和着床能力等多个方面影响ART的成功率。精子DNA损伤的因素主要包括氧化应激、年龄、激素以及环境等。同时,卵母细胞对精子DNA损伤的修复能力也至关重要。目前临床和实验室主要通过药物干预、优化精子选择来降低精子DNA碎片指数(DNA fragmentation index,DFI),从而改善ART的临床治疗结局。综述精子DNA损伤的研究现状,分析其对ART治疗结局的影响,并提出了可能的改进精液制备方法。

在ICSI时应用SperMagic对无精子症患者辅助生殖结局的影响

目的:探究在卵胞浆内单精子显微注射(ICSI)时应用SperMagic对无精子症患者的临床效果。方法:选取2019年1月至2021年1月佛山市妇幼保健院收治的200例移植周期的无精子症患者,遵循随机数字表法将纳入对象随机分为观察组及对照组,各100例。其中对照组患者精子处理后行ICSI,观察组患者在精子处理后添加精子激动剂SperMagic。比较两组患者精子活力以及辅助生殖结局等相关指标差异。结果:精子在经相应处理后,观察组患者的精子活力为(92.6±3.6)%,与对照组精子活力(67.6±11.4)%相比,差异具有统计学意义(P<0.05)。与对照组相比,观察组患者的受精率、优胚率、种植率、妊娠率及流产率等辅助生殖结局指标显著更优,差异具有统计学意义(P<0.05)。结论:在ICSI时加入精子激动剂SperMagic,可显著提高精子活力,改善妊娠结局。

Tales of the Tail and Sperm Head Aches --Changing concepts on the prognostic significance of sperm pathologies affecting the head, neck and tail

This article presents an update on the variable prognostic significance of different sperm pathologies in patients with severe male factor infertility due to morphology and motility disorders. Severe asthenozoospermia is one of the leading causes of male infertility as spermatozoa cannot reach the oocyte and/or penetrate normally. Identifying structural causes of sperm immotility was of great concern before the advent of intracytoplasmic sperm injection （ICSI）, because immotility was the limiting factor in the treatment of these patients. In these cases, in vitro methods are used to identify live spermatozoa or stimulate sperm motility to avoid selection of non-viable cells. With these advances, fertilization and pregnancy results have improved dramatically. The identification of genetic phenotypes in asthenozoospermia is important to adequately inform patients of treatment outcomes and risks. The one sperm characteristic that seriously affects fertility prognosis is teratozoospermia, primarily sperm head and neck anomalies. Defects of chromatin condensation and acrosomal hypoplasia are the two most common abnormalities in severe teratozoospermia. The introduction of microscopic methods to select spermatozoa and the development of new ones to evaluate sperm quality before ICSI will assure that ultrastructural identification ofsperm pathologies will not only be of academic interest, but will also be an essential tool to inform treatment choice. Herein, we review the differential roles played by sperm components in normal fertilization and early embryo development and explore how assisted reproductive technologies have modified our concepts on the prognostic significance of sperm pathologies affecting the head, neck, mid-piece and tail.

Comparison of sperm retrieval and reproductive outcome in azoospermic men with testicular failure and obstructive azoospermia treated for infertility

We assessed the rates of sperm retrieval and intracytoplasmic sperm injection outcomes, including the neonatal profile of infants conceived, in men with testicular failure. Three-hundred and sixty-five men with testicular failure who underwent micro-dissection testicular sperm extraction were included in this study. We compared their outcomes with 40 men with testicular failure who used donor sperm for injections due to failed retrieval, and 146 men with obstructive azoospermia who underwent percutaneous sperm retrieval. The retrieval rate in testicular failure was 41.4%, and the results were lower than the obstructed azoospermia （100%; adjusted odds ratio： 0.033; 95% Ch 0.007-0.164; P 〈 0.001）. Live birth rates after sperm injections were lower in men with testicular failure （19.9%） compared with donor sperm （37.5%; adjusted OR： 0.377 （95% Ch 0.233-0.609, P 〈 0.001）） and obstructive azoospermia （34.2%; adjusted OR： 0.403 （95% CI： 0.241-0.676, P= 0.001）. Newborn parameters of infants conceived were not significantly different among the groups. We concluded that the chances of obtaining sperm on retrieval and achieving a live birth after intracytoplasmic sperm injection （ICSI） are reduced in men with testicular failure. The profile of infants conceived after sperm injection does not seem to be negatively affected by testicular failure.

No relationship between biopsy sites near the main testicular vessels or rete testis and successful sperm retrieval using conventional or microdissection biopsies in 220 non-obstructive azoospermic men

In 220 consecutive patients with non-obstructive azoospermia, sperm retrieval was attempted by a combination of conventional and microdissection testicular sperm extraction （TESE）. For sperm retrieval, 2-3 conventional biopsies were performed followed by a microdissection TESE in cases of negative conventional biopsies. During the surgery, the vasculature of the testis was assessed using the operative microscope, and the location of positive biopsies was registered in relation to the blood supply. The overall sperm retrieval rate was 58.2%. From the initial conventional biopsies, sperm could be retrieved in 46.8% of the patients. With microdissection TESE, sperm could be retrieved from an additional 11.4% of the patients. The further use of microdissection TESE improved the sperm retrieval rate significantly （P=0.017）. No significant accumulation of positive biopsies was found towards the rete testis or the main testicular vessels.

Application of seminal germ cell morphology and semen biochemistry in the diagnosis and management of azoospermic subjects

Aim: To evaluate whether the study of seminal germ cell morphology (SGCM) and semen biochemistry could befruitfully utilized for the diagnosis and management of azoospermic subjects. Methods: In the semen, mature andimmature germ cells are contributed by the testes, 70% of glycerylphosphoryl choline (GPC) by the epididymis, fruc-tose mostly or solely by the seminal vesicles and acid phosphate (ACP) by the prostate. In 16 normal volunteers, 12vasectomized subjects and 186 azoospennic subjects, these parameters have been studied and the data have been ana-lyzed. Results: Both mature and immature germ cells are absent in the semen of vasectomized subjects as well as inobstructive azoospennia; GPC level is also significantly decreased in both these groups. In cases with non-obstructiveazoospermia immature germ cells are present and seminal GPC, ACP and fructose levels are normal. The diagnosis ofobstructive and non-obstructive azoospermia based on these parameters correlated well with 'correct' testicular biopsyfindings. In some cases of azoospermia due to hypospermatogenesis or spermatogenic developmental arrest, the SGCMstudies were very helpful in objectively monitoring the response of the germinal tissue to specific treaunents. Conclu-sion: SGCM and semen biochemical parameters are very valuable non-invasive markers for differentiating obstructivefrom non-obstructive azoospermia. The SGCM findings serve as a dependable non-invasive testicular marker with highpredictive value. (Asian J Androl 2001 Mar; 3: 55-62)