Shanghai Key Laboratory of Reproductive Medicine

Shanghai, China

Shanghai Key Laboratory of Reproductive Medicine

Shanghai, China

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Hou D.,Shanghai JiaoTong University | Hou D.,Shanghai Key Laboratory of Reproductive Medicine | Zhou X.,University of Idaho | Zhong X.,University of Idaho | And 8 more authors.
Fertility and Sterility | Year: 2013

Objective To explore potential causes of male infertility by determining the composition and structure of commensal bacterial communities in seminal fluids. Design Microscopy of Gram-stained semen samples and classification of 16S rRNA gene sequences to determine the species composition of semen bacterial communities. Setting Clinical andrology laboratory and academic research laboratories. Patient(s) Nineteen sperm donors and 58 infertility patients. Intervention(s) None. Main Outcome Measure(s) Classification of 16S rRNA gene sequences, clustering of seminal microbial communities, and multiple statistical tests. Result(s) High numbers of diverse kinds of bacteria were present in most samples of both sperm donors and infertility patients. The bacterial communities varied widely among subjects, but they could be clustered into six groups based on similarities in composition and the rank abundances of taxa. Overall, there were no significant differences between sperm donors and infertility patients. However, multiple statistical tests showed a significant negative association between sperm quality and the presence of Anaerococcus. The results also indicated that many of the bacterial taxa identified in semen also occur in the vaginal communities of some women, especially those with bacterial vaginosis, which suggests that heterosexual sex partners may share bacteria. Conclusion(s) Diverse kinds of bacteria were present in the human semen, but there were no significant differences between sperm donors and infertility patients. The presence of Anaerococcus might be a biomarker for low sperm quality. © 2013 by American Society for Reproductive Medicine.


Mei X.-X.,Shanghai JiaoTong University | Wang J.,Shanghai JiaoTong University | Wu J.,Shanghai JiaoTong University | Wu J.,Ningxia Medical University | Wu J.,Shanghai Key Laboratory of Reproductive Medicine
Asian Journal of Andrology | Year: 2015

Spermatogonial stem cells (SSCs), the stem cells responsible for male fertility, are one of a small number of cells with the abilities of both self-renewal and generation of large numbers of haploid cells. Technology improvements, most importantly, transplantation assays and in vitro culture systems have greatly expanded our understanding of SSC self-renewal and differentiation. Many important molecules crucial for the balance between self-renewal and differentiation have been recently identified although the exact mechanism(s) remain largely undefined. In this review, we give a brief introduction to SSCs, and then focus on extrinsic and intrinsic factors controlling SSCs self-renewal and differentiation. © 2015 AJA, SIMM & SJTU. All rights reserved 1008-682X.


Zhang C.,Shanghai JiaoTong University | Wu J.,Shanghai JiaoTong University | Wu J.,Ningxia Medical University | Wu J.,Shanghai Key Laboratory of Reproductive Medicine
Molecular Human Reproduction | Year: 2016

Study hypothesis:We investigated whether DEAD-box polypeptide 4 (DDX4) positive cells from post-natal ovaries of germline lineage reporter mice can be isolated based on endogenously expressed fluorescent proteins and used to establish a cell line for producing offspring. study finding: DDX4-positive cells frompost-natal ovaries of germline lineage reporter mice can be isolated and used to establish a cell line for producing offspring. what is known already: In recent years, female germline stem cells (FGSCs) have been isolated from the ovaries of post-natal mice by magnetic-activated cell sorting or fluorescence-activated cell sorting (FACS) relying on an antibody against DDX4. However, whether DDX4- positive cells frompost-natal ovaries of germline lineage reporter mice can be established without using an antibody, as well as a cell line established for producing offspring, remains unknown. study design, samples/materials, methods: To obtain the expected offspring (Ddx4-Cre;mT/mGmice), Ddx4-Cre micewere crossed withmT/mGmice. In the ovaries of Ddx4-Cre;mT/mGmice, germ cellswere destined to express enhanced green fluorescent protein (EGFP) while somatic cells still express tandem dimer Tomato (tdTomato). Therefore, the germ cells could be clearly distinguished from somatic cells by fluorescent proteins. Then, we investigated the pattern of fluorescent cells in the ovaries of 21-day-old Ddx4-Cre;mT/mG mice under a fluorescent microscope.Germ cells were sorted by FACS without using antibody and used to establish a FGSC line. The FGSC line was analyzed by DDX4 immunostaining, Edu (5-ethynyl-2'-deoxyuridine) labeling, and RT-PCR for germ cell markers. Finally, the physiological function of the FGSC line was examined by transplanting FGSCs into the ovaries of sterilized recipients and subsequent mating. main results and the role of chance: Firstly, we have successfully isolated FGSCs from the ovaries of 21-day-old Ddx4- Cre;mT/mG mice based on endogenously expressed fluorescent proteins. FACS was used to separate the cells and 2.3% of all viable cells was EGFP-positive germ cells. Subsequently, a FGSC line was established that was doubly positive for DDX4 immunostaining and Edu labeling. The mRNA expression of several germ cell markers in this cell line, such as Ddx4, Deleted in azoospermia-like (Dazl), B lymphocyte-induced maturation protein-1 (Blimp1), Stella and Fragilis,was detected. Lastly, the FGSClinewas proven to be functional under physiological conditions, as offspringwere produced after transplanting FGSCs into ovaries of sterilized recipients and a subsequent mating. limitations, reasons for caution: The molecular mechanisms of proliferation and differentiation of FGSCs in vivo and in vitro still need to be elucidated. wider implications of the findings: Our results confirm that DDX4-positive cells can be separated from post-natal mouse ovaries and used to establish cell lines that are functional in producing off spring, and provide further evidence for the existence of post-natal FGSCs in mammals. The Ddx4-Cre;mT/mG mouse strain is an ideal model for the isolation, characterization and propagation of FGSCs and is a useful tool for fully elucidating the molecular mechanisms of proliferation and differentiation of FGSCs in vivo and in vitro. large scale data: None. study funding and competing interest(s): This work was supported by National Basic Research Program of China (2013CB967401) and the National Nature Science Foundation of China (81370675, 81200472 and 81421061). The authors declare no competing interests. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.


Guo Y.,Shanghai JiaoTong University | Hai Y.,Shanghai JiaoTong University | Gong Y.,Shanghai JiaoTong University | Li Z.,Shanghai JiaoTong University | And 3 more authors.
Journal of Cellular Physiology | Year: 2014

Spermatogenesis is a special process by which spermatogonial stem cells (SSCs) divide and differentiate to male gametes called mature spermatozoa. SSCs are the unique cells because they are adult stem cells that transmit genetic information to subsequent generations. Accumulating evidence has demonstrated that SSCs can be reprogrammed to acquire pluripotency to become embryonic stem-like cells that differentiate into all cell lineages of the three germ layers, highlighting potential important applications of SSCs for regenerative medicine. Recent studies from peers and us have made great achievements on the characterization, isolation, and culture of mouse and human SSCs, which could lead to better understanding the biology of SSCs and the applications of SSCs in both reproductive and regenerative medicine. In this review, we first compared the cell identity and biochemical phenotypes between mouse SSCs and human SSCs. Notably, the cell types of mouse and human SSCs are distinct, and human SSCs share some but not all phenotypes with mouse SSCs. The approaches for isolating SSCs as well as short- and long-term culture of mouse SSCs and short-period culture of human SSCs were also discussed. We further addressed the new advances on the self-renewal of SSCs with an aim to establish the long-term culture of human SSCs which has not yet been achieved. © 2013 Wiley Periodicals, Inc.


Wu J.,Shanghai JiaoTong University | Wu J.,Shanghai Key Laboratory of Reproductive Medicine | Bao J.,Shanghai JiaoTong University | Bao J.,Shanghai Key Laboratory of Reproductive Medicine | And 6 more authors.
BMC Developmental Biology | Year: 2011

Background: There have been increasing attentions on the role of small RNAs, especially microRNAs in post-transcriptional gene regulation during spermatogenesis. MicroRNA-184 (miR-184) has been shown to be mainly expressed in the testis and brain, and that its expression levels are by far the highest in the testis. However, the role of miR-184 in mammalian spermatogenesis remains unclear. Results: In this study, we demonstrated that miR-184 levels were increased during mouse postnatal testis development. Specifically, miR-184 expression was restricted to the germ cells from spermatogonia to round spermatids. Overexpression of miR-184 promoted the proliferation of a germ cell line, GC-1spg. Moreover, miR-184 downregulated nuclear receptor corepressor 2 (Ncor2) by targeting its 3' untranslated region through inhibiting NCOR2 protein translation. Conclusions: MiR-184 may be involved in the post-transcription regulation of mRNAs such as Ncor2 in mammalian spermatogenesis. © 2011 Wu et al; licensee BioMed Central Ltd.


Hai Y.,Shanghai JiaoTong University | Hou J.,Shanghai JiaoTong University | Liu Y.,Shanghai JiaoTong University | Yang H.,Shanghai JiaoTong University | And 4 more authors.
Seminars in Cell and Developmental Biology | Year: 2014

Spermatogenesis is a complex process by which spermatogonial stem cells (SSCs) self-renew and differentiate into spermatozoa under the elaborate coordination of testicular microenvironment, namely, niche. Sertoli cells, which locate around male germ cells, are the most critical component of the niche. Significant progress has recently been made by peers and us on uncovering the effects of Sertoli cells on regulating fate determinations of SSCs. Here we addressed the roles and regulation of Sertoli cells in normal and abnormal spermatogenesis. Specifically, we summarized the biological characteristics of Sertoli cells, and we emphasized the roles of Sertoli cells in mediating the self-renewal, differentiation, apoptosis, de-differentiation, and trans-differentiation of SSCs. The association between abnormal function of Sertoli cells and impaired spermatogenesis was discussed. Finally, we highlighted several issues to be addressed for further investigation on the effects and mechanisms of Sertoli cells in spermatogenesis. Since Sertoli cells are the key supportive cells for SSCs and they are very receptive to modification, a better understanding of the roles and regulation of Sertoli cells in SSC biology and spermatogenesis would make it feasible to identify novel targets for gene therapy of male infertility as well as seek more efficient and safer strategies for male contraception. © 2014 Elsevier Ltd.


Wang L.,Shanghai JiaoTong University | Wang L.,Shanghai Key Laboratory of Reproductive Medicine | Xu C.,Shanghai JiaoTong University | Xu C.,Shanghai Key Laboratory of Reproductive Medicine
Reproduction | Year: 2015

microRNAs (miRNAs) are a class of small endogenous RNAs, 19-25 nucleotides in size, which play a role in the regulation of gene expression at transcriptional and post-transcriptional levels. Spermatogenesis is a complex process through which spermatogonial stem cells (SSCs) proliferate and differentiate into mature spermatozoa. A large number of miRNAs are abundantly expressed in spermatogenic cells. Growing evidence supports the essential role of miRNA regulation in normal spermatogenesis and male fertility and cumulative research has shown that this form of regulation contributes to the etiology of testicular germ cell tumors (TGCTs). In this review, we addressed recent advancements of miRNA expression profiles in testis and focused on the regulatory functions of miRNA in the process of SSC renewal, spermatogonial mitosis, spermatocyte meiosis, spermiogenesis, and the occurrence of TGCTs. © 2015 Society for Reproduction and Fertility.


Zhang Z.,Shanghai JiaoTong University | Gong Y.,Shanghai JiaoTong University | Guo Y.,Shanghai JiaoTong University | Hai Y.,Shanghai JiaoTong University | And 9 more authors.
Cell Communication and Signaling | Year: 2013

Background: Severe shortage of liver donors and hepatocytes highlights urgent requirement of extra-liver and stem cell source of hepatocytes for treating liver-related diseases. Here we hypothesized that spermatogonial stem cells (SSCs) can directly transdifferentiate to hepatic stem-like cells capable of differentiating into mature hepatocyte-like cells in vitro without an intervening pluripotent state. Results: SSCs first changed into hepatic stem-like cells since they resembled hepatic oval cells in morphology and expressed Ck8, Ck18, Ck7, Ck19, OV6, and albumin. Importantly, they co-expressed CK8 and CK19 but not ES cell markers. Hepatic stem-like cells derived from SSCs could differentiate into small hepatocytes based upon their morphological features and expression of numerous hepatic cell markers but lacking of bile epithelial cell hallmarks. Small hepatocytes were further coaxed to differentiate into mature hepatocyte-like cells, as identified by their morphological traits and strong expression of Ck8, Ck18, Cyp7a1, Hnf3b, Alb, Tat, Ttr, albumin, and CYP1A2 but not Ck7 or CK19. Notably, these differentiated cells acquired functional attributes of hepatocyte-like cells because they secreted albumin, synthesized urea, and uptake and released indocyanine green. Moreover, phosphorylation of ERK1/2 and Smad2/3 rather than Akt was activated in hepatic stem cells and mature hepatocytes. Additionally, cyclin A, cyclin B and cyclin E transcripts and proteins but not cyclin D1 or CDK1 and CDK2 transcripts or proteins were reduced in mature hepatocyte-like cells or hepatic stem-like cells derived from SSCs compared to SSCs. Conclusions: SSCs can transdifferentiate to hepatic stem-like cells capable of differentiating into cells with morphological, phenotypic and functional characteristics of mature hepatocytes via the activation of ERK1/2 and Smad2/3 signaling pathways and the inactivation of cyclin A, cyclin B and cyclin E. This study thus provides an invaluable source of mature hepatocytes for treating liver-related diseases and drug toxicity screening and offers novel insights into mechanisms of liver development and cell reprogramming. © 2013 Zhang et al.; licensee BioMed Central Ltd.


Xia X.,Shanghai JiaoTong University | Xia X.,Shanghai Key Laboratory of Reproductive Medicine | Cai H.,Shanghai JiaoTong University | Cai H.,Shanghai Key Laboratory of Reproductive Medicine | And 4 more authors.
PLoS ONE | Year: 2012

In the previous study, we unraveled the unique "erasure strategy" during the mouse spermiogenesis. Chromatin associated proteins sequentially disassociated from the spermatid chromosome, which led to the termination of transcription in elongating spermatids. By this process, a relatively naïve paternal chromatin was generated, which might be essential for the zygotic development. We supposed the regulation of histone acetylation played an important role throughout this "erasure" process. In order to verify this hypothesis, we treated mouse spermatids in vitro by histone acetylase (HAT) inhibitor Curcumin. Our results showed an inhibiting effect of Curcumin on the growth of germ cell line in a dose-dependent manner. Accordingly, the apoptosis of primary haploid spermtids was increased by Curcumin treatment. As expected, the acetylated histone level was downregulated. Furthermore, we found the transcription in spermatids ceased in advance, the dynamics of chromatin associated factors was disturbed by Curcumin treatment. The regulation of histone acetylation should be one of the core reprogramming mechanisms during the spermiogenesis. The reproductive toxicity of Curcumin needs to be thoroughly investigated, which is crucial for its further clinical application. © 2012 Xia et al.


Wang Y.,Chongqing Medical University | Du Y.,Nanjing University | Shen B.,Nanjing Medical University | Zhou X.,Chongqing Medical University | And 11 more authors.
Scientific Reports | Year: 2015

Co-injection of zygotes with Cas9 mRNA and sgRNA has been proven to be an efficient gene-editing strategy for genome modification of different species. Genetic engineering in pigs holds a great promise in biomedical research. By co-injection of one-cell stage embryos with Cas9 mRNA and Npc1l1 sgRNA, we achieved precise Npc1l1 targeting in Chinese Bama miniature pigs at the efficiency as high as 100%. Meanwhile, we carefully analyzed the Npc1l1 sgRNA:Cas9-mediated on- and off-target mutations in various somatic tissues and ovaries, and demonstrated that injection of zygotes with Cas9 mRNA and sgRNA is an efficient and reliable approach for generation of gene-modified pigs.

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