In vitro production of haploid cells after coculture of CD49f+ with Sertoli cells from testicular sperm extraction in nonobstructive azoospermic patients

Slides:

Advertisements

Similar presentations

In vitro differentiation of germ cells from nonobstructive azoospermic patients using three-dimensional culture in a collagen gel matrix Jae-Ho Lee,

Advertisements

Expression of activated HER2 in human testes

Rong Hu, Ph. D. , Fei-miao Wang, M. D. , Liang Yu, Ph. D. , Yan Luo, M

Study of nucleation status in the second cell cycle of human embryo and its impact on implantation rate Jesus Aguilar, Ph.D., Irene Rubio, Ph.D., Elkin.

Differential expression of G-protein-coupled estrogen receptor-30 in human myometrial and uterine leiomyoma smooth muscle Ruijuan Tian, M.Sc., Zengyong.

Human CD133+ bone marrow-derived stem cells promote endometrial proliferation in a murine model of Asherman syndrome Irene Cervelló, Ph.D., Claudia Gil-Sanchis,

Human spermatogonial stem cells display limited proliferation in vitro under mouse spermatogonial stem cell culture conditions Jose V. Medrano, Ph.D.,

Testicular expression of synaptonemal complex protein 3 (SYCP3) messenger ribonucleic acid in 110 patients with nonobstructive azoospermia Mahmoud Aarabi,

Adenomyosis does not affect implantation, but is associated with miscarriage in patients undergoing oocyte donation José A. Martínez-Conejero, Ph.D.,

Increased expression of macrophage colony–stimulating factor and its receptor in patients with endometriosis Nicole M. Budrys, M.D., M.P.H., Hareesh.

Coenzyme Q–dependent mitochondrial respiratory chain activity in granulosa cells is reduced with aging Assaf Ben-Meir, M.D., Shlomi Yahalomi, M.D., Brit.

Outdoor air pollution and sperm quality

Meiotic arrest at the midpachytene stage in a patient with complete azoospermia factor b deletion of the Y chromosome Jeanne Perrin, M.D., Catherine.

Cell selection by selective matrix adhesion is not sufficiently efficient for complete malignant cell depletion from contaminated human testicular cell.

Composition of single-step media used for human embryo culture

Peritubular cells may modulate Leydig cell–mediated testosterone production through a nonclassic pathway Jibanananda Mishra, Ph.D., Mukesh Gautam, M.Sc.,

Expression of forkhead transcription factors in human granulosa cells

Comparative evaluation of different in vitro systems that stimulate germ cell differentiation in human embryonic stem cells Mark Richards, Ph.D., Chui-Yee.

Follicular fluid and mural granulosa cells microRNA profiles vary in in vitro fertilization patients depending on their age and oocyte maturation stage

Can an educational DVD improve the acceptability of elective single embryo transfer? A randomized controlled study Nicole Hope, M.B., B.S., Hon., Luk.

High serum FSH levels in men with nonobstructive azoospermia does not affect success of microdissection testicular sperm extraction Ranjith Ramasamy,

Outcomes of microdissection testicular sperm extraction in men with nonobstructive azoospermia due to maturation arrest Aaron M. Bernie, M.D., M.P.H.,

Azoospermia: is sample centrifugation indicated

Deleted in Azoospermia-Like (DAZL) gene–expressing cells in human amniotic fluid: a new source for germ cells research? Konstantinos Stefanidis, M.D.,

Regine Gaetje, M. D. , Uwe Holtrich, Ph. D. , Thomas Karn, Ph. D

In vitro effects of phthalate esters in human myometrial and leiomyoma cells and increased urinary level of phthalate metabolite in women with uterine.

Comparison of two methods of in vivo gene transfer by electroporation

How old is too old? Challenges faced by clinicians concerning age cutoffs for patients undergoing in vitro fertilization Robert L. Klitzman, M.D. Fertility.

Trends in clinical reproductive medicine research: 10 years of growth

Genetic variants in the ETV5 gene in fertile and infertile men with nonobstructive azoospermia associated with Sertoli cell–only syndrome Moira K. O'Bryan,

Ramadan Saleh, M. D. , Reda Z. Mahfouz, M. D. , Ashok Agarwal, Ph. D

Estrogenic regulation of testicular expression of stem cell factor and c-kit: implications in germ cell survival and male fertility Sara Correia, M.S.,

Beneficial value of testicular sperm extraction-AgarCyto in addition to the standard testicular biopsy for diagnosis of testicular germ cell tumors in.

Klinefelter's syndrome with unilateral absence of vas deferens

Sperm DNA fragmentation levels in testicular sperm samples from azoospermic males as assessed by the sperm chromatin dispersion (SCD) test Marcos Meseguer,

Evaluation of a human ovarian follicle isolation technique to obtain disease-free follicle suspensions before safely grafting to cancer patients Michelle.

Effect of the male factor on the clinical outcome of intracytoplasmic sperm injection combined with preimplantation aneuploidy testing: observational.

Heat shock factor Y chromosome (HSFY) mRNA level predicts the presence of retrievable testicular sperm in men with nonobstructive azoospermia Peter J.

Cysteine-rich secretory protein 1 in seminal plasma: potential biomarker for the distinction between obstructive and nonobstructive azoospermia Christine.

Adenovirus-mediated expression of cyclooxygenase-2 antisense reverse abnormal genetic profile of human adhesion fibroblasts Ghassan M. Saed, Ph.D., Ayman.

Integrative DNA methylation and gene expression analysis identifies discoidin domain receptor 1 association with idiopathic nonobstructive azoospermia

Anna Kékesi, Edit Erdei, M. D. , Ph. D. , Miklós Török, M. D. , Ph. D

Application of three-dimensional fluorescence in situ hybridization to human preimplantation genetic diagnosis Li-Ying Yan, Ph.D., Jie Qiao, M.D., Yuan.

Alternate indications for varicocele repair: non-obstructive azoospermia, pain, androgen deficiency and progressive testicular dysfunction Peter N. Schlegel,

Ana Cervero, Ph. D. , Francisco Domínguez, Ph. D

Volume 3, Issue 4, Pages (October 2014)

Leucine-rich repeat–containing G-protein–coupled receptor 5–positive cells in the endometrial stem cell niche Irene Cervelló, Ph.D., Claudia Gil-Sanchis,

Cumulative newborn rates increase with the total number of transferred embryos according to an analysis of 15,792 ovum donation cycles Nicolás Garrido,

Evaluation of diagnostic testis biopsy and the repetition of testicular sperm extraction surgeries in infertility patients Alayman Hussein, M.B.B.C.H.,

Aline Bozec, Ph. D. , Souheila Amara, Ph. D. , Basma Guarmit, M. Sc

Lysed cell removal promotes frozen–thawed embryo development

Human papillomavirus found in sperm head of young adult males affects the progressive motility Carlo Foresta, M.D., Andrea Garolla, M.D., Daniela Zuccarello,

Microarray analysis in sperm from fertile and infertile men without basic sperm analysis abnormalities reveals a significantly different transcriptome

Epidermal growth factor–containing fibulin-like extracellular matrix protein 1 expression and regulation in uterine leiomyoma Erica E. Marsh, M.D., M.S.C.I.,

Roberto Ensenat-Waser, Ph. D. , Antonio Pellicer, M. D. , Ph. D

Impact of different patterns of sperm chromosomal abnormalities on the chromosomal constitution of preimplantation embryos Lorena Rodrigo, B.Sc., Vanessa.

Human myometrium and leiomyomas express gonadotropin-releasing hormone 2 and gonadotropin-releasing hormone 2 receptor Jason D. Parker, D.O., Minnie.

Dimensional assessment of X-bearing and Y-bearing haploid and disomic human sperm with the use of fluorescence in situ hybridization and objective morphometry

Higher PDCD4 expression is associated with obesity, insulin resistance, lipid metabolism disorders, and granulosa cell apoptosis in polycystic ovary syndrome

Reciprocal translocation t(7;16)(q21.2;p13.3) in an infertile man

Association, prevalence, and clearance of human papillomavirus and antisperm antibodies in infected semen samples from infertile patients Andrea Garolla,

Insulin growth factors may be implicated in human sperm capacitation

MicroRNA: key gene expression regulators

Comparison of microdissection testicular sperm extraction, conventional testicular sperm extraction, and testicular sperm aspiration for nonobstructive.

Giovanna Mantovani, M. D. , Mario Mancini, M. D. , Giacomo Gazzano, M

Genetic evaluation procedures at sperm banks in the United States

Fei Sun, M. D. , Ph. D. , Paul Turek, M. D. , Calvin Greene, M. D

In vitro sildenafil citrate use as a sperm motility stimulant

Are urinary bisphenol A levels in men related to semen quality and embryo development after medically assisted reproduction? Jure Knez, M.D., Roman Kranvogl,

Claire E. Beyer, B. Sc. , Bettina Kayler, B. M. Sc

Presentation transcript:

In vitro production of haploid cells after coculture of CD49f+ with Sertoli cells from testicular sperm extraction in nonobstructive azoospermic patients Marcia Riboldi, B.Sc., Carmen Rubio, Ph.D., Antonio Pellicer, M.D., Ph.D., Manuel Gil-Salom, Ph.D., Carlos Simón, M.D., Ph.D. Fertility and Sterility Volume 98, Issue 3, Pages 580-590.e4 (September 2012) DOI: 10.1016/j.fertnstert.2012.05.039 Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 1 (A) Flow cytometry isolation of CD49f cells. (B) (I) Real-time polymerase chain reaction (RT-PCR) for THY-1, GAF, C-KIT, DAZL, VASA, STELLA, PIWILL2, BOULE, DCM1, SCP3, PRM1, TNP2, and Blimp1 in CD49f+ cells. (IIa) RT-PCR for positive control. (IIb) RT-PCR for negative control. (C) Immunocytochemistry in CD49f positive cells: (I) CD49f, (II) dapi, and (III) merge; (IV) THY-1, (V) dapi, and (VI) merge; (VII) GPR125, (VIII) dapi, and (IX) merge. Fertility and Sterility 2012 98, 580-590.e4DOI: (10.1016/j.fertnstert.2012.05.039) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 2 (A) Flow cytometry for Nile red-positive staining in testicular cells. (B) (I) Lipid droplet by oil red in Sertoli cells (magnification: ×10). (II) Lipid droplet by oil red in Sertoli cells (magnification: ×40). (C) (I) Lipid droplet by electron microscopy in Sertoli cells. (II) Lipid droplet by electron microscopy in Sertoli cells. (III) Tight junctions by electron microscopy in Sertoli cells. (IV) Tight junctions by electron microscopy in Sertoli cells. (D) (I) Real-time polymerase chain reaction (RT-PCR) for GAF, SCF, GATA-4, and WT-1 in Sertoli cells. (IIa) RT-PCR for positive control. (IIb) RT-PCR for negative control. (E) Immunocytochemistry in Sertoli cells for (I) GATA-4, (II) dapi, (III) merge; (IV) vimentin, (V) dapi, and (VI) merge. (F) Sertoli cells express red fluorescent protein (RFP). Fertility and Sterility 2012 98, 580-590.e4DOI: (10.1016/j.fertnstert.2012.05.039) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 3 (A) Fluorescence in situ hybridization (FISH) in CD49f+ cells: (I) dapi, (II) chromosome 18, (III) chromosome X, and (IV) chromosome Y. (B) FISH in Sertoli cells: (I) dapi, (II) chromosome 18, (III) chromosome X, and (IV) chromosome Y. (C) Immunocytochemistry for meiosis in CD49f+ cells (staining: negative for SCP3, CREST, and MLH1; positive for DAPI). (D) Immunocytochemistry for meiosis in Sertoli cells red fluorescent protein (RFP) (staining: negative for SCP3, CREST, and MLH1; positive for DAPI). Fertility and Sterility 2012 98, 580-590.e4DOI: (10.1016/j.fertnstert.2012.05.039) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 4 (A) Immunocytochemistry for meiosis in nonobstructive azoospermic (NOA) coculture on day 3. (I–IV) Positive staining for SCP3 and CREST. (B) Immunocytochemistry for meiosis in NOA coculture on day 5. (I) Positive for SCP3 and CREST. (II) Negative for SCP3, CREST, and MLH1. (III) Positive for SCP3. (IV) Positive for SCP3 and CREST. (C) Immunocytochemistry for meiosis in NOA coculture on day 7. (I) Positive for SCP3. (II) Negative for SCP3, CREST, and MLH1. (III) Positive for SCP3. (IV) Positive for SCP3. (D) FISH for the four chromosomes in NOA by CD49f+ cells: (I) dapi, (II) chromosome 13, (III) chromosome 21, (IV) chromosome 18, and (V) chromosome X. Fertility and Sterility 2012 98, 580-590.e4DOI: (10.1016/j.fertnstert.2012.05.039) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 Mean estimate of positive staining for meiosis markers (SCP3, CREST, MLH1). “Before coculture” is the CD49f+ and Sertoli red fluorescent protein (RFP) control before the start of coculture. “Percentage coculture” is the time course of the cells in coculture (day 3 to day 10). D = diplotene (positive staining for SCP3, CREST, and MLH1); L = leptotene (positive staining for SCP3); P = pachytene (positive staining for SCP3, CREST, and sometimes MLH1); NA = not available (data not shown); NOA = nonobstructive azoospermia; OA = obstructive azoospermia; Z = zygotene (positive staining for SCP3 and sometimes CREST). Fertility and Sterility 2012 98, 580-590.e4DOI: (10.1016/j.fertnstert.2012.05.039) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions