Interaction between thyroid hormones and gonadotropin inhibitory hormone in ex vivo culture of zebrafish testis: an approach to study multifactorial control of spermatogenesis

ABSTRACTReproduction is under multifactorial control of neurohormones, pituitary gonadotropins, as well as a number of gonadal hormones including sex steroids and growth factors. Gonadotropin-inhibitory hormone (Gnih), a novel RFamide neuropeptide, was shown to be involved in the control of pituitary gonadotropin production, as well as being involved as a paracrine factor in the regulation of gonadal function. In this context, recent studies have demonstrated that Gnih inhibited gonadotropin-induced spermatogenesis in the zebrafish testicular explants. Thyroid hormones are known to interact with the reproductive axis, and are, in particular, involved in the regulation of testicular function. Based on this background, we investigated the interaction between Gnih and thyroid hormones in the control of zebrafish spermatogenesis. To this end, zebrafish adult males were treated with the goitrogen methimazole (1mM for 21 days) in order to generate a hypothyroid model organism. Subsequently, a factorial design using an ex vivo testis culture system in combination with histomorphometrical and FACScan cell cycle analyses were adopted. Our results showed that methimazole treatment affected both basal and gonadotropin-induced spermatogenesis, in particular, meiosis and spermiogenesis. Moreover, the goitrogen treatment nullified the inhibitory actions of Gnih on the gonadotropin-induced spermatogenesis, specifically in the haploid cell population. We have demonstrated that thyroid hormones interaction with gonadotropin and Gnih are important components for the regulation of zebrafish spermatogenesis. The results provide a support for the hypothesis that thyroid hormones are important contributors in multifactorial control of spermatogenesis in zebrafish.

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Phagocytosis of Liposome Particles by Rat Splenic Immature Monocytes Makes Them Transiently and Highly Immunosuppressive In Ex Vivo Culture Conditions

Journal of Pharmacology and Experimental Therapeutics ◽

10.1124/jpet.110.172510 ◽

2011 ◽

Vol 337 (1) ◽

pp. 42-49 ◽

Cited By ~ 8

Author(s):

Daisuke Takahashi ◽

Hiroshi Azuma ◽

Hiromi Sakai ◽

Keitaro Sou ◽

Daiko Wakita ◽

...

Keyword(s):

Ex Vivo ◽

Culture Conditions ◽

Ex Vivo Culture

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Mimicking of Blood Flow Results in a Distinct Functional Phenotype in Human Non-Adherent Classical Monocytes

Biology ◽

10.3390/biology10080748 ◽

2021 ◽

Vol 10 (8) ◽

pp. 748

Author(s):

Elisa Wirthgen ◽

Melanie Hornschuh ◽

Ida Maria Wrobel ◽

Christian Manteuffel ◽

Jan Däbritz

Keyword(s):

Blood Flow ◽

Shear Flow ◽

Ex Vivo ◽

Culture Conditions ◽

Inflammatory Signaling ◽

Gm Csf ◽

Beneficial Effects ◽

Migratory Capacity ◽

Ex Vivo Culture ◽

Static Conditions

Ex vivo culture conditions during the manufacturing process impact the therapeutic effect of cell-based products. Mimicking blood flow during ex vivo culture of monocytes has beneficial effects by preserving their migratory ability. However, the effects of shear flow on the inflammatory response have not been studied so far. Hence, the present study investigates the effects of shear flow on both blood-derived naïve and activated monocytes. The activation of monocytes was experimentally induced by granulocyte-macrophage colony-stimulating factor (GM-CSF), which acts as a pro-survival and growth factor on monocytes with a potential role in inflammation. Monocytes were cultured under dynamic (=shear flow) or static conditions while preventing monocytes' adherence by using cell-repellent surfaces to avoid adhesion-induced differentiation. After cultivation (40 h), cell size, viability, and cytokine secretion were evaluated, and the cells were further applied to functional tests on their migratory capacity, adherence, and metabolic activity. Our results demonstrate that the application of shear flow resulted in a decreased pro-inflammatory signaling concurrent with increased secretion of the anti-inflammatory cytokine IL-10 and increased migratory capacity. These features may improve the efficacy of monocyte-based therapeutic products as both the unwanted inflammatory signaling in blood circulation and the loss of migratory ability will be prevented.

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Stem cell integrins: Implications for ex-vivo culture and cellular therapies

Stem Cell Research ◽

10.1016/j.scr.2010.09.005 ◽

2011 ◽

Vol 6 (1) ◽

pp. 1-12 ◽

Cited By ~ 110

Author(s):

Andrew B.J. Prowse ◽

Fenny Chong ◽

Peter P. Gray ◽

Trent P. Munro

Keyword(s):

Stem Cell ◽

Ex Vivo ◽

Cellular Therapies ◽

Ex Vivo Culture

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Ex vivo culture of Fancc-/- stem/progenitor cells predisposes cells to undergo apoptosis, and surviving stem/progenitor cells display cytogenetic abnormalities and an increased risk of malignancy

Blood ◽

10.1182/blood-2004-06-2483 ◽

2005 ◽

Vol 105 (9) ◽

pp. 3465-3471 ◽

Cited By ~ 48

Author(s):

Xiaxin Li ◽

Michelle M. Le Beau ◽

Samantha Ciccone ◽

Feng-Chun Yang ◽

Brian Freie ◽

...

Keyword(s):

Progenitor Cells ◽

Genome Stability ◽

Bone Marrow Cells ◽

Ex Vivo ◽

Acquired Resistance ◽

Intrinsic Defects ◽

Cytogenetic Abnormalities ◽

Increased Risk ◽

Ex Vivo Culture ◽

The Impact

AbstractCurrent strategies for genetic therapy using Moloney retroviruses require ex vivo manipulation of hematopoietic cells to facilitate stable integration of the transgene. While many studies have evaluated the impact of ex vivo culture on normal murine and human stem/progenitor cells, the cellular consequences of ex vivo manipulation of stem cells with intrinsic defects in genome stability are incompletely understood. Here we show that ex vivo culture of Fancc-/- bone marrow cells results in a time-dependent increase in apoptosis of primitive Fancc-/- progenitor cells in conditions that promote the proliferation of wild-type stem/progenitor cells. Further, recipients reconstituted with the surviving Fancc-/- cells have a high incidence of cytogenetic abnormalities and myeloid malignancies that are associated with an acquired resistance to tumor necrosis factor α (TNF-α). Collectively, these data indicate that the intrinsic defects in the genomic stability of Fancc-/- stem/progenitor cells provide a selective pressure for cells that are resistant to apoptosis and have a propensity for the evolution to clonal hematopoiesis and malignancy. These studies could have implications for the design of genetic therapies for treatment of Fanconi anemia and potentially other genetic diseases with intrinsic defects in genome stability.

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The presence of the carboxy-terminal fragment of fibronectin allows maintenance of non-human primate long-term hematopoietic repopulating cells during extended ex vivo culture and transduction

Experimental Hematology ◽

10.1016/j.exphem.2003.10.003 ◽

2004 ◽

Vol 32 (2) ◽

pp. 163-170 ◽

Cited By ~ 15

Author(s):

S Sellers

Keyword(s):

Ex Vivo ◽

Terminal Fragment ◽

Ex Vivo Culture ◽

Carboxy Terminal ◽

Human Primate

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Establishing an ex vivo culture system for normal pancreatic tissue

Pancreatology ◽

10.1016/j.pan.2014.05.789 ◽

2014 ◽

Vol 14 (3) ◽

pp. S120-S121

Author(s):

Carlos Fernández Moro ◽

Sougat Misra ◽

Soledad Pouso ◽

Marita Wallenberg ◽

Rainer Heuchel ◽

...

Keyword(s):

Culture System ◽

Ex Vivo ◽

Pancreatic Tissue ◽

Normal Pancreatic Tissue ◽

Ex Vivo Culture

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Ex vivo culture of human CD34+ cord blood cells with thrombopoietin (TPO) accelerates platelet engraftment in a NOD/SCID mouse model

Experimental Hematology ◽

10.1016/j.exphem.2006.04.009 ◽

2006 ◽

Vol 34 (7) ◽

pp. 943-950 ◽

Cited By ~ 27

Author(s):

Yvette van Hensbergen ◽

Laurus F. Schipper ◽

Anneke Brand ◽

Manon C. Slot ◽

Mick Welling ◽

...

Keyword(s):

Mouse Model ◽

Cord Blood ◽

Blood Cells ◽

Scid Mouse ◽

Ex Vivo ◽

Human Cd34 ◽

Scid Mouse Model ◽

Cord Blood Cells ◽

Ex Vivo Culture

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Assessment of Long-Term in vitro Multiplied Human Wharton’s Jelly-Derived Mesenchymal Stem Cells prior to Their Use in Clinical Administration

Cells Tissues Organs ◽

10.1159/000517423 ◽

2021 ◽

pp. 1-11

Author(s):

Urvi Panwar ◽

Kanchan Mishra ◽

Parizad Patel ◽

Sumit Bharadva ◽

Salil Vaniawala ◽

...

Keyword(s):

Telomere Length ◽

Replicative Senescence ◽

Ex Vivo ◽

Wharton’S Jelly ◽

Cellular Aging ◽

Human Umbilical Cord ◽

Surface Markers ◽

Wharton's Jelly ◽

Ex Vivo Culture

The quantity of mesenchymal stem/stromal cells (MSCs) required for a particular therapy demands their subsequent expansion through ex vivo culture. During in vitro multiplication, they undergo replicative senescence which may alter their genetic stability. Therefore, this study was aimed to analyze cellular, molecular, and chromosomal alterations in Wharton’s jelly-derived MSCs (WJ-MSCs) during their in vitro sequential passages, where WJ-MSCs were sequentially passaged up to P14 and cells were evaluated at an interval of P2, P6, P10, and P14. They were examined for their morphology, tumorigenicity, surface markers, stemness markers, DNA damage, chromosomal aberration, and telomere length. We have processed five full-term delivered human umbilical cord samples to obtain WJ-MSCs. Morphological appearance observed at initial stages was small fine spindle-shaped WJ-MSCs which were transformed to flat, long, and broader cells in later passages. The cell proliferation rate was gradually decreased after the 10th passage. WJ-MSCs have expressed stemness markers OCT-4 and NANOG, while they showed high expression of positive surface markers CD90 and CD105 and lower expression of CD34 and CD45. They were non-tumorigenic with slow cellular aging during subsequent passages. There was no chromosomal abnormality up to the 14th passage, while increase in comet score and decrease in telomere length were observed in later passages. Hence, our study suggests that early and middle passaged (less than P10) WJ-MSCs are good candidates for clinical administration for treatment.

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