In vitro culture of spermatogonial stem cells （SSCs） has generally been performed using two-dimensional （2D） culture systems; however, such cultures have not led to the development of complete spermatogenesis. It ...In vitro culture of spermatogonial stem cells （SSCs） has generally been performed using two-dimensional （2D） culture systems; however, such cultures have not led to the development of complete spermatogenesis. It seems that 2D systems do not replicate optimal conditions of the seminiferous tubules （including those generated by the SSC niche） and necessary for spermatogenesis. Recently, one of our laboratories has been able to induce proliferation and differentiation of mouse testicular germ cells to meiotic and postmeiotic stages including generation of sperm in a 3D soft agar culture system （SACS） and a 3D methylcellulose culture system （MCS）. It was suggested that SACS and MCS form a special 3D microenvironment that mimics germ cell niche formation in the seminiferous tubules, and thus permits mouse spermatogenesis in vitro. In this review, we （1） provide a brief overview of the differences in spermatogenesis in rodents and primates, （2） summarize data related to attempts to generate sperm in vitro, （3） report for the first time formation of colonies/clusters of cells and differentiation of meiotic （expression of CREM-1） and postmeiotic （expression of acrosin） germ cells from undifferentiated spermatogonia isolated from the testis of prepubertal rhesus monkeys and cultured in SACS and MCS, and （4） indicate research needed to optimize 3D systems for in vitro primate spermatogenesis and for possible future application to man.展开更多
Background:Despite the high prevalence of strongyloidiasis in the Laotian population,Laotian hospitals still lack diagnostic capacity to appropriately diagnose Strongyloides stercoralis infections.This cross-sectional...Background:Despite the high prevalence of strongyloidiasis in the Laotian population,Laotian hospitals still lack diagnostic capacity to appropriately diagnose Strongyloides stercoralis infections.This cross-sectional hospital-based study was conducted to assess the prevalence of Strongyloides stercoralis infection among hospitalized patients treated at Mahosot Hospital,the primary reference hospital of Lao People’s Democratic Republic(Lao PDR),and to validate feasible methods for diagnosing S.stercoralis infection at hospital’s laboratory.Methods:Between September and December 2018,stool samples of 104 inpatients were investigated for S.stercoralis infection by wet smear,Baermann technique,Koga Agar plate culture(KAPC),and real-time detection polymerase chain reaction(RTD-PCR)at the Infectious Diseases Ward of the Mahosot Hospital in Vientiane.The sensitivity,the specificity,the negative predictive value(NPV)of each diagnostic test,as well as their combination(s)was calculated using a composite reference standard(CRS).The correlation of the different test methods was assessed by chi-square or Fisher’s exact test.Cohen’s kappa coefficient was used to assess the diagnostic agreement of the different test methods.Results:The overall prevalence of S.stercoralis infections among the study population was 33.4%.The cumulative infection prevalence statistically significantly increased from the lowest age group of 40 years and below(22.4%),to the medium(40.0%)and to the oldest age group of 61 year and above(72.7%)(P=0.003).The cumulative infection prevalence of CRS was considerably higher in male(40.4%)compared to female patients(28.1%),but not statistically different(P=0.184).The diagnostic sensitivity of Baermann technique,KAPC,RTD-PCR,and the combination of Baermann technique and KAPC were 60.0,60.0,74.3,and 77.1%,respectively.Only 13 patients(37.1%)of the total 35 S.stercoralis patients diagnosed with any technique had a simultaneously positive diagnostic test with Baermann,KAPC and RTD-PCR.Conclusions:We identified Baermann technique and KAPC to be currently the most feasible and implementable standard methods for diagnosing S.stercoralis at a hospital setting such as Mahosot Hospital and provincial and district hospitals in Lao PDR and other low-and middle income countries in Southeast Asia.Trial registration:This study was approved by the National Ethics Committee for Health Research in Lao PDR(reference no.083/NECHR)and by the Ethics Committee Northwest and Central Switzerland(reference no.2018–00594).展开更多
文摘In vitro culture of spermatogonial stem cells （SSCs） has generally been performed using two-dimensional （2D） culture systems; however, such cultures have not led to the development of complete spermatogenesis. It seems that 2D systems do not replicate optimal conditions of the seminiferous tubules （including those generated by the SSC niche） and necessary for spermatogenesis. Recently, one of our laboratories has been able to induce proliferation and differentiation of mouse testicular germ cells to meiotic and postmeiotic stages including generation of sperm in a 3D soft agar culture system （SACS） and a 3D methylcellulose culture system （MCS）. It was suggested that SACS and MCS form a special 3D microenvironment that mimics germ cell niche formation in the seminiferous tubules, and thus permits mouse spermatogenesis in vitro. In this review, we （1） provide a brief overview of the differences in spermatogenesis in rodents and primates, （2） summarize data related to attempts to generate sperm in vitro, （3） report for the first time formation of colonies/clusters of cells and differentiation of meiotic （expression of CREM-1） and postmeiotic （expression of acrosin） germ cells from undifferentiated spermatogonia isolated from the testis of prepubertal rhesus monkeys and cultured in SACS and MCS, and （4） indicate research needed to optimize 3D systems for in vitro primate spermatogenesis and for possible future application to man.
文摘Background:Despite the high prevalence of strongyloidiasis in the Laotian population,Laotian hospitals still lack diagnostic capacity to appropriately diagnose Strongyloides stercoralis infections.This cross-sectional hospital-based study was conducted to assess the prevalence of Strongyloides stercoralis infection among hospitalized patients treated at Mahosot Hospital,the primary reference hospital of Lao People’s Democratic Republic(Lao PDR),and to validate feasible methods for diagnosing S.stercoralis infection at hospital’s laboratory.Methods:Between September and December 2018,stool samples of 104 inpatients were investigated for S.stercoralis infection by wet smear,Baermann technique,Koga Agar plate culture(KAPC),and real-time detection polymerase chain reaction(RTD-PCR)at the Infectious Diseases Ward of the Mahosot Hospital in Vientiane.The sensitivity,the specificity,the negative predictive value(NPV)of each diagnostic test,as well as their combination(s)was calculated using a composite reference standard(CRS).The correlation of the different test methods was assessed by chi-square or Fisher’s exact test.Cohen’s kappa coefficient was used to assess the diagnostic agreement of the different test methods.Results:The overall prevalence of S.stercoralis infections among the study population was 33.4%.The cumulative infection prevalence statistically significantly increased from the lowest age group of 40 years and below(22.4%),to the medium(40.0%)and to the oldest age group of 61 year and above(72.7%)(P=0.003).The cumulative infection prevalence of CRS was considerably higher in male(40.4%)compared to female patients(28.1%),but not statistically different(P=0.184).The diagnostic sensitivity of Baermann technique,KAPC,RTD-PCR,and the combination of Baermann technique and KAPC were 60.0,60.0,74.3,and 77.1%,respectively.Only 13 patients(37.1%)of the total 35 S.stercoralis patients diagnosed with any technique had a simultaneously positive diagnostic test with Baermann,KAPC and RTD-PCR.Conclusions:We identified Baermann technique and KAPC to be currently the most feasible and implementable standard methods for diagnosing S.stercoralis at a hospital setting such as Mahosot Hospital and provincial and district hospitals in Lao PDR and other low-and middle income countries in Southeast Asia.Trial registration:This study was approved by the National Ethics Committee for Health Research in Lao PDR(reference no.083/NECHR)and by the Ethics Committee Northwest and Central Switzerland(reference no.2018–00594).
