scholarly journals Long-Term Oocyte-Like Cell Development in Cultures Derived from Neonatal Marmoset Monkey Ovary

2016 ◽  
Vol 2016 ◽  
pp. 1-17 ◽  
Author(s):  
Bentolhoda Fereydouni ◽  
Gabriela Salinas-Riester ◽  
Michael Heistermann ◽  
Ralf Dressel ◽  
Lucia Lewerich ◽  
...  
Keyword(s):  
Stem Cell ◽  
Germ Cell ◽  
Germ Cells ◽  
Cell Biology ◽  
Culture System ◽  
Cultured Cells ◽  
Embryonic Stem ◽  
Cell Development ◽  
Stem Cell Markers ◽  
Marmoset Monkey

We use the common marmoset monkey (Callithrix jacchus) as a preclinical nonhuman primate model to study reproductive and stem cell biology. The neonatal marmoset monkey ovary contains numerous primitive premeiotic germ cells (oogonia) expressing pluripotent stem cell markers including OCT4A (POU5F1). This is a peculiarity compared to neonatal human and rodent ovaries. Here, we aimed at culturing marmoset oogonia from neonatal ovaries. We established a culture system being stable for more than 20 passages and 5 months. Importantly, comparative transcriptome analysis of the cultured cells with neonatal ovary, embryonic stem cells, and fibroblasts revealed a lack of germ cell and pluripotency genes indicating the complete loss of oogonia upon initiation of the culture. From passage 4 onwards, however, the cultured cells produced large spherical, free-floating cells resembling oocyte-like cells (OLCs). OLCs strongly expressed several germ cell genes and may derive from the ovarian surface epithelium. In summary, our novel primate ovarian cell culture initially lacked detectable germ cells but then produced OLCs over a long period of time. This culture system may allow a deeper analysis of early phases of female primate germ cell development and—after significant refinement—possibly also the production of monkey oocytes.

scholarly journals Survival Motor Neuron Protein Participates in Mouse Germ Cell Development and Spermatogonium Maintenance

2020 ◽  
Vol 21 (3) ◽  
pp. 794 ◽  
Author(s):  
Wei-Fang Chang ◽  
Jie Xu ◽  
Tzu-Ying Lin ◽  
Jing Hsu ◽  
Hsiu-Mei Hsieh-Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Motor Neuron ◽  
Cell Biology ◽  
Critical Role ◽  
Embryonic Stem ◽  
Cell Development ◽  
Survival Motor Neuron ◽  
Specific Marker ◽  
Germ Cell Development

The defective human survival motor neuron 1 (SMN1) gene leads to spinal muscular atrophy (SMA), the most common genetic cause of infant mortality. We previously reported that loss of SMN results in rapid differentiation of Drosophila germline stem cells and mouse embryonic stem cells (ESCs), indicating that SMN also plays important roles in germ cell development and stem cell biology. Here, we show that in healthy mice, SMN is highly expressed in the gonadal tissues, prepubertal spermatogonia, and adult spermatocytes, whereas low SMN expression is found in differentiated spermatid and sperm. In SMA-like mice, the growth of testis tissues is retarded, accompanied with gamete development abnormalities and loss of the spermatogonia-specific marker. Consistently, knockdown of Smn1 in spermatogonial stem cells (SSCs) leads to a compromised regeneration capacity in vitro and in vivo in transplantation experiments. In SMA-like mice, apoptosis and accumulation of the R-loop structure were significantly elevated, indicating that SMN plays a critical role in the survival of male germ cells. The present work demonstrates that SMN, in addition to its critical roles in neuronal development, participates in mouse germ cell and spermatogonium maintenance.

scholarly journals A novel embryonic stem cell line derived from the common marmoset monkey (Callithrix jacchus) exhibiting germ cell-like characteristics

2009 ◽  
Vol 24 (6) ◽  
pp. 1359-1372 ◽  
Author(s):  
T. Muller ◽  
G. Fleischmann ◽  
K. Eildermann ◽  
K. Matz-Rensing ◽  
P. A. Horn ◽  
...  
Keyword(s):  
Stem Cell ◽  
Germ Cell ◽  
Embryonic Stem Cell ◽  
Cell Line ◽  
Embryonic Stem Cell Line ◽  
Embryonic Stem ◽  
Common Marmoset ◽  
Stem Cell Line ◽  
Marmoset Monkey ◽  
The Common

scholarly journals 50 years of spermatogenesis: Sertoli cells and their interactions with germ cells

2018 ◽  
Vol 99 (1) ◽  
pp. 87-100 ◽  
Author(s):  
Michael D Griswold
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Cultured Cells ◽  
Cell Types ◽  
Cell Development ◽  
Specific Cell ◽  
Germ Cell Development ◽  
Cell Niche ◽  
Transgenic Technologies

Abstract The complex morphology of the Sertoli cells and their interactions with germ cells has been a focus of investigators since they were first described by Enrico Sertoli. In the past 50 years, information on Sertoli cells has transcended morphology alone to become increasingly more focused on molecular questions. The goal of investigators has been to understand the role of the Sertoli cells in spermatogenesis and to apply that information to problems relating to male fertility. Sertoli cells are unique in that they are a nondividing cell population that is active for the reproductive lifetime of the animal and cyclically change morphology and gene expression. The numerous and distinctive junctional complexes and membrane specializations made by Sertoli cells provide a scaffold and environment for germ cell development. The increased focus of investigators on the molecular components and putative functions of testicular cells has resulted primarily from procedures that isolate specific cell types from the testicular milieu. Products of Sertoli cells that influence germ cell development and vice versa have been characterized from cultured cells and from the application of transgenic technologies. Germ cell transplantation has shown that the Sertoli cells respond to cues from germ cells with regard to developmental timing and has furthered a focus on spermatogenic stem cells and the stem cell niche. Very basic and universal features of spermatogenesis such as the cycle of the seminiferous epithelium and the spermatogenic wave are initiated by Sertoli cells and maintained by Sertoli-germ cell cooperation.

scholarly journals 213.Culture of mouse male germ cells for genetic manipulations

2004 ◽  
Vol 16 (9) ◽  
pp. 213
Author(s):  
C. M. Itman ◽  
B. Barakat ◽  
K. Loveland
Keyword(s):  
Gene Transfer ◽  
Germ Cell ◽  
Germ Cells ◽  
Cell Biology ◽  
Culture System ◽  
Striking Difference ◽  
Embryonal Carcinoma Cell ◽  
Cells In Culture ◽  
New Genes

The ability to maintain germ cells in culture offers the opportunity to manipulate their differentiation and to deliver new genes through the germline. However the study of male germ cell biology is hindered by the lack of an in vitro culture system that supports germ cell maintenance and differentiation, and the lack of efficient gene transfer technologies. To address this, we developed a short term (1-6 day) in vitro germ cell-Sertoli cell co-culture system using testes of 6 day old mice, in which the only germ cell type present is the spermatogonium. To establish culture conditions favourable for maintaining germ cells in culture, we compared different substrates (plastic, laminin, poly-L-lysine, Matrigel) and cell culture additives (insulin/transferrin, retinoic acid, lactic acid, pyruvic acid). Using immunocytochemistry, we observed that germ cell numbers varied depending on the substrate, and was greatly improved on laminin or Matrigel. The culture additives tested had no effect. We noted a striking difference in germ cell growth depending on the mouse strain used, with C57BlXCBA F1 cross > Swiss > BalbC >> C57Bl/6J. To investigate gene transfer into germ cells, constructs were generated using promoters identified as exhibiting germ cell-specific expression in transgenic animals (EF1-α, Oct4, Stra8) with EGFP and lacz reporter genes. From this baseline, we have started gene transfer experiments. Initial transfection experiments were performed in mouse testis cell lines (GC1, GC2, TM3, TM4) and the P19 mouse embryonal carcinoma cell line. Several commercial transfection agents and a non-commercial product (1) were used, as was electroporation, however these did not yield reporter expression in germ cells. As these techniques require proliferating cells, we are now undertaking a study with the addition of growth factors in culture (BMP4, activin, GDNF, FSH) to enhance germ cell proliferation and therefore improve transfection efficiencies. We will also employ lentiviral transduction, as this is reportedly 100% efficient and independent of proliferation. (1) Celebi et al. (2002) Mol. Reprod. Dev. 62,477–482.

scholarly journals Disorganization of the germ cell pool leads to primary ovarian insufficiency

Reproduction ◽  
2017 ◽  
Vol 153 (6) ◽  
pp. R205-R213 ◽  
Author(s):  
Ikko Kawashima ◽  
Kazuhiro Kawamura
Keyword(s):  
Germ Cell ◽  
Cell Biology ◽  
Embryonic Stem ◽  
Primordial Follicle ◽  
Cell Development ◽  
Primary Ovarian Insufficiency ◽  
Ovarian Insufficiency ◽  
Germ Cell Development ◽  
Female Germ Cell

The mammalian ovary is an organ that controls female germ cell development, storing them and releasing mature oocytes for transporting to the oviduct. During the fetal stage, female germ cells change from a proliferative state to meiosis before forming follicles with the potential for the growth of surrounding somatic cells. Understanding of molecular and physiological bases of germ cell development in the fetal ovary contributed not only to the elucidation of genetic disorders in primary ovarian insufficiency (POI), but also to the advancement of novel treatments for patients with POI. Accumulating evidence indicates that mutations inNOBOX,DAZLandFIGLAgenes are associated with POI. In addition, cell biology studies revealed the important roles of these genes as essential translational factors for germ cell development. Recent insights into the role of the PI3K (phosphatidylinositol 3-kinase)-Akt signaling pathway in primordial follicle activation allowed the development of a new infertility treatment, IVA (in vitroactivation), leading to successful pregnancy/delivery in POI patients. Furthermore, elucidation of genetic dynamics underlying female germ cell development could allow regeneration of oocytes from ES (embryonic stem)/iPS (induced pluripotent stem) cells in mammals. The purpose of this review is to summarize basic findings related to female germ cell development and potential clinical implications, especially focusing on POI etiologies. We also summarize evolving new POI therapies based on IVA as well as oocyte regeneration.

scholarly journals Spontaneously Formed Spheroids from Mouse Compact Bone-Derived Cells Retain Highly Potent Stem Cells with Enhanced Differentiation Capability

2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
Kai Chen ◽  
Xianqi Li ◽  
Ni Li ◽  
Hongwei Dong ◽  
Yiming Zhang ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cultured Cells ◽  
Embryonic Stem ◽  
Compact Bone ◽  
Enzymatic Digestion ◽  
Stem Cell Markers ◽  
Water Contact ◽  
Cell Markers ◽  
Polymerase Chain ◽  
Embryonic Stem Cell Markers

The results from our recent study showed the presence of two distinct spheroid-forming mechanisms, i.e., spontaneous and mechanical. In this study, we focused on the spontaneously formed spheroids, and the character of spontaneously formed spheroids from mouse compact bone-derived cells (CBDCs) was explored. Cells from (C57BL/6J) mouse leg bones were isolated, and compact bone-derived cells were cultured after enzymatic digestion. Spontaneous spheroid formation was achieved on a culture plate with specific water contact angle as reported. The expression levels of embryonic stem cell markers were analyzed using immunofluorescence and quantitative reverse transcription polymerase chain reaction. Then, the cells from spheroids were induced into osteogenic and neurogenic lineages. The spontaneously formed spheroids from CBDCs were positive for ES cell markers such as SSEA1, Sox2, Oct4, and Nanog. Additionally, the expressions of fucosyltransferase 4/FUT4 (SSEA1), Sox2, and Nanog were significantly higher than those in monolayer cultured cells. The gene expression of mesenchymal stem cell markers was almost identical in both spheroids and monolayer-cultured cells, but the expression of Sca-1 was higher in spheroids. Spheroid-derived cells showed significantly higher osteogenic and neurogenic marker expression than monolayer-cultured cells after induction. Spontaneously formed spheroids expressed stem cell markers and showed enhanced osteogenic and neurogenic differentiation capabilities than cells from the conventional monolayer culture, which supports the superior stemness.

scholarly journals EGFR confers exquisite specificity of Wnt9a-Fzd9b signaling in hematopoietic stem cell development

2018 ◽  
Author(s):  
Stephanie Grainger ◽  
Nicole Nguyen ◽  
Jenna Richter ◽  
Jordan Setayesh ◽  
Brianna Lonquich ◽  
...  
Keyword(s):  
Stem Cell ◽  
Wnt Signaling ◽  
Hematopoietic Stem Cell ◽  
Cell Biology ◽  
Embryonic Stem ◽  
Cell Development ◽  
List Type ◽  
Hematopoietic Stem

SummaryThe mechanisms of Wnt-Frizzled (Fzd) signaling selectivity and their biological implications remain unclear. We demonstrate for the first time that the epidermal growth factor receptor (EGFR) is required as a co-factor for Wnt signaling. Using genetic studies in zebrafish, paired within vitrocell biology and biochemistry, we have determined that Fzd9b signals specifically with Wnt9ain vivoandin vitroto elicit β-catenin dependent Wnt signals that regulate hematopoietic stem and progenitor cell (HSPC) development in the dorsal aorta. This requirement is conserved in the derivation of HSPCs from human embryonic stem cells. Wnt9a-Fzd9b specificity requires two intracellular domains in Fzd9b, which interact with EGFR as a required co-factor to promote signal transduction. EGFR phosphorylates one tyrosine residue on Fzd9b, a requirement for the Wnt signal. These findings indicate that Wnt signaling interactions can be exquisitely specific and inform protocols for derivation of HSPCsin vitro.HighlightsAnin vitrosignaling screen identifies Fzd9b as a Wnt9a-specific receptor.Fzd9b and Wnt9a regulate hematopoietic stem cell development as a cognate pair.WNT9A and FZD9 are required for HSPC derivation from human pluripotent cellsin vitro.EGFR confers specificity to Wnt9a-Fzd9b signaling in zebrafish and human cells.

scholarly journals Cyclosporin A and FGF signaling support the proliferation/survival of mouse primordial germ cell-like cells in vitro†

2020 ◽  
Author(s):  
Hiroshi Ohta ◽  
Yukihiro Yabuta ◽  
Kazuki Kurimoto ◽  
Tomonori Nakamura ◽  
Yusuke Murase ◽  
...  
Keyword(s):  
Germ Cell ◽  
Cyclosporin A ◽  
Germ Cells ◽  
Culture System ◽  
Ex Vivo ◽  
Primordial Germ Cell ◽  
Embryonic Stem ◽  
Chemical Screening

Abstract Primordial germ cells (PGCs) are the founding population of the germ cell lineage that undergo a multistep process to generate spermatozoa or oocytes. Establishing an appropriate culture system for PGCs is a key challenge in reproductive biology. By a chemical screening using mouse PGC-like cells (mPGCLCs), which were induced from mouse embryonic stem cells, we reported previously that forskolin and rolipram synergistically enhanced the proliferation/survival of mPGCLCs with an average expansion rate of ~20-fold. In the present study, we evaluated other chemicals or cytokines to see whether they would improve the current mPGCLC culture system. Among the chemicals and cytokines examined, in the presence of forskolin and rolipram, cyclosporin A (CsA) and fibroblast growth factors (FGFs: FGF2 and FGF10) effectively enhanced the expansion of mPGCLCs in vitro (~50-fold on average). During the expansion by CsA or FGFs, mPGCLCs comprehensively erased their DNA methylation to acquire a profile equivalent to that of gonadal germ cells in vivo, while maintaining their highly motile phenotype as well as their transcriptional properties as sexually uncommitted PGCs. Importantly, these mPGCLCs robustly contributed to spermatogenesis and produced fertile offspring. Furthermore, mouse PGCs (mPGCs) cultured with CsA ex vivo showed transcriptomes and DNA methylomes similar to those of cultured mPGCLCs. The improved culture system for mPGCLCs/mPGCs would be instructive for addressing key questions in PGC biology, including the mechanisms for germ cell migration, epigenetic reprogramming, and sex determination of the germline.

The expression of mesenchymal and embryonic stem cell markers by human limbal Stromal cells

2011 ◽  
Author(s):  
Moon Nian Lim ◽  
Umapathy Thiageswari ◽  
Othman Ainoon ◽  
P. J. N. Baharuddin ◽  
R. A. Jamal ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Stromal Cells ◽  
Embryonic Stem ◽  
Stem Cell Markers ◽  
Cell Markers ◽  
Embryonic Stem Cell Markers

scholarly journals PRL3 induces polypoid giant cancer cells eliminated by PRL3-zumab to reduce tumor relapse

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Min Thura ◽  
Zu Ye ◽  
Abdul Qader Al-Aidaroos ◽  
Qiancheng Xiong ◽  
Jun Yi Ong ◽  
...  
Keyword(s):  
Dna Damage ◽  
Stem Cell ◽  
Cancer Cells ◽  
Embryonic Stem ◽  
Genotoxic Stress ◽  
Stem Cell Markers ◽  
Tumor Removal ◽  
Primary Tumors ◽  
Tumor Relapse ◽  
Dna Damage Signaling

AbstractPRL3, a unique oncotarget, is specifically overexpressed in 80.6% of cancers. In 2003, we reported that PRL3 promotes cell migration, invasion, and metastasis. Herein, firstly, we show that PRL3 induces Polyploid Giant Cancer Cells (PGCCs) formation. PGCCs constitute stem cell-like pools to facilitate cell survival, chemo-resistance, and tumor relapse. The correlations between PRL3 overexpression and PGCCs attributes raised possibilities that PRL3 could be involved in PGCCs formation. Secondly, we show that PRL3+ PGCCs co-express the embryonic stem cell markers SOX2 and OCT4 and arise mainly due to incomplete cytokinesis despite extensive DNA damage. Thirdly, we reveal that PRL3+ PGCCs tolerate prolonged chemotherapy-induced genotoxic stress via suppression of the pro-apoptotic ATM DNA damage-signaling pathway. Fourthly, we demonstrated PRL3-zumab, a First-in-Class humanized antibody drug against PRL3 oncotarget, could reduce tumor relapse in ‘tumor removal’ animal model. Finally, we confirmed that PGCCs were enriched in relapse tumors versus primary tumors. PRL3-zumab has been approved for Phase 2 clinical trials in Singapore, US, and China to block all solid tumors. This study further showed PRL3-zumab could potentially serve an ‘Adjuvant Immunotherapy’ after tumor removal surgery to eliminate PRL3+ PGCC stem-like cells, preventing metastasis and relapse.

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