scholarly journals Treatment with retinoic acid and lens epithelial cell-conditioned medium in vitro directed the differentiation of pluripotent stem cells towards corneal endothelial cell-like cells

2014 ◽  
Vol 9 (2) ◽  
pp. 351-360 ◽  
Author(s):  
PING CHEN ◽  
JUN-ZHAO CHEN ◽  
CHUN-YI SHAO ◽  
CHUAN-YIN LI ◽  
YI-DAN ZHANG ◽  
...  
Keyword(s):  
Stem Cells ◽  
Retinoic Acid ◽  
Endothelial Cell ◽  
Epithelial Cell ◽  
Conditioned Medium ◽  
Pluripotent Stem Cells ◽  
Lens Epithelial Cell ◽  
Corneal Endothelial Cell ◽  
Cell Conditioned Medium

scholarly journals Review: corneal endothelial cell derivation methods from ES/iPS cells

2019 ◽  
Vol 39 (1) ◽  
Author(s):  
Shin Hatou ◽  
Shigeto Shimmura
Keyword(s):  
Endothelial Cell ◽  
Conditioned Medium ◽  
Corneal Transplantation ◽  
Ips Cells ◽  
Lens Epithelial Cell ◽  
Corneal Endothelial Cell ◽  
Bullous Keratopathy ◽  
Corneal Endothelial Dysfunction ◽  
Fetal Bovine ◽  
Proof Of Efficacy

Abstract Globally, approximately 12.7 million people are awaiting a transplantation, while only 185,000 cases of corneal transplantation are performed in a year. Corneal endothelial dysfunction (bullous keratopathy) due to Fuchs’ corneal endothelial dystrophy, or insults associated with intraocular surgeries, shared half of all indications for corneal transplantation. Regenerative therapy for corneal endothelium independent of eye bank eyes has great importance to solve the large supply-demand mismatching in corneal transplantation and reduce the number of worldwide corneal blindness. If corneal endothelial cells could be derived from ES or iPS cells, these stem cells would be the ideal cell source for cell therapy treatment of bullous keratopathy. Four representative corneal endothelial cell derivation methods were reviewed. Components in earlier methods included lens epithelial cell-conditioned medium or fetal bovine serum, but the methods have been improved and materials have been chemically more defined over the years. Conditioned medium or serum is replaced to recombinant proteins and small molecule compounds. These improvements enabled to open the corneal endothelial developmental mechanisms, in which epithelial-mesenchymal and mesenchymal-endothelial transition by TGF beta, BMP, and Wnt signaling have important roles. The protocols are gradually approaching clinical application; however, proof of efficacy and safety of the cells by adequate animal models are the challenges for the future.

scholarly journals Transcriptome-based molecular staging of human stem cell-derived retinal organoids uncovers accelerated photoreceptor differentiation by 9-cis retinal

2019 ◽  
Author(s):  
Koray D. Kaya ◽  
Holly Y. Chen ◽  
Matthew J. Brooks ◽  
Ryan A. Kelley ◽  
Hiroko Shimada ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Retinoic Acid ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Mitochondrial Morphology ◽  
Rod Photoreceptor ◽  
Molecular Staging ◽  
Organoid Cultures

ABSTRACTRetinal organoids generated from human pluripotent stem cells exhibit considerable variability in temporal dynamics of differentiation. To assess the maturity of neural retina in vitro, we performed transcriptome analyses of developing organoids from human embryonic and induced pluripotent stem cell lines. We show that the developmental variability in organoids was reflected in gene expression profiles and could be evaluated by molecular staging with the human fetal and adult retinal transcriptome data. We also demonstrated that addition of 9-cis retinal, instead of widely-used all-trans retinoic acid, accelerated rod photoreceptor differentiation in organoid cultures, with higher rhodopsin expression and more mature mitochondrial morphology evident by day 120. Our studies thus provide an objective transcriptome-based modality for determining the differentiation state of retinal organoids, which should facilitate disease modeling and evaluation of therapies in vitro.Summary StatementThree-dimensional organoids derived from human pluripotent stem cells have been extensively applied for investigating organogenesis, modeling diseases and development of therapies. However, substantial variations within organoids pose challenges for comparison among different cultures and studies. We generated transcriptomes of multiple distinct retinal organoids and compared these to human fetal and adult retina gene profiles for molecular staging of differentiation state of the cultures. Our analysis revealed the advantage of using 9-cis retinal, instead of the widely-used all-trans retinoic acid, in facilitating rod photoreceptor differentiation. Thus, a transcriptome-based comparison can provide an objective method to uncover the maturity of organoid cultures across different lines and in various study platforms.

Retinoic acid and/or progesterone differentiate mouse induced pluripotent stem cells into male germ cells in vitro

2019 ◽  
Vol 121 (3) ◽  
pp. 2159-2169
Author(s):  
Javad Amini Mahabadi ◽  
Abolfazl Aazami Tameh ◽  
Sayyed Alireza Talaei ◽  
Mohammad Karimian ◽  
Tahereh Rahiminia ◽  
...  
Keyword(s):  
Stem Cells ◽  
Retinoic Acid ◽  
Induced Pluripotent Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Male Germ Cells ◽  
Induced Pluripotent

Endothelium-dependent ANF secretion in vitro

1992 ◽  
Vol 263 (4) ◽  
pp. H1071-H1077 ◽  
Author(s):  
R. A. Lew ◽  
A. J. Baertschi
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Conditioned Medium ◽  
Superoxide Anion ◽  
Heat Stable ◽  
Guanylate Cyclase Activator ◽  
Stimulatory Factor ◽  
Cell Conditioned Medium ◽  
Stimulation Of

Coculture of endothelial cells with atrial cells (R. A. Lew and A. J. Baertschi. Biochem. Biophys. Res. Commun. 163: 701-709, 1989) increased atrial natriuretic factor (ANF) release to 205 +/- 15% (n = 33 experiments) of basal secretion (2.02 +/- 0.33 ng/ml). Stimulation of ANF release by endothelial cells was significantly reduced (P < 0.05) by addition of the calcium channel antagonist nicardipine (Nic, 100 nM; by 69 +/- 4%), the guanylate cyclase activator sodium nitroprusside (SNP, 1 microM; by 97 +/- 27%), or acetylcholine (ACh, 10 microM; by 55 +/- 13%). Endothelial cell-conditioned medium elicited a 62 +/- 10% (n = 10) increase in ANF release. Rat and porcine endothelin (0.1-100 nM) each elicited a dose-dependent increase in ANF release [up to 84 +/- 14% (n = 18) over baseline]. The activity of conditioned medium was not affected by heat or trypsin treatment, but was significantly reduced by addition of Nic or SNP and was attenuated by ACh. Stimulation of ANF by 1 nM synthetic rat or porcine endothelin was also unaffected by heat or trypsin but was significantly reduced by Nic, SNP, and ACh. Addition of endothelin-specific antiserum abolished the ANF stimulatory activity of endothelial cell-conditioned medium. Neither inhibition of superoxide anion by superoxide dismutase nor inhibition of endothelium-derived nitric oxide production by NG-monomethyl-L-arginine affected the ANF release from coculture. Thus endothelial cells release a heat-stable, diffusible ANF stimulatory factor, which is not endothelium-derived relaxing factor or superoxide anion but is biologically and immunologically similar to endothelin.

A bronchial epithelial cell-derived factor in asthma that promotes eosinophil activation and survival as GM-CSF

1991 ◽  
Vol 260 (6) ◽  
pp. L530-L538 ◽  
Author(s):  
M. Soloperto ◽  
V. L. Mattoso ◽  
A. Fasoli ◽  
S. Mattoli
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Conditioned Medium ◽  
Calcium Ionophore ◽  
Bronchial Epithelial Cells ◽  
Eosinophil Infiltration ◽  
Bronchial Epithelial ◽  
Gm Csf ◽  
Cell Conditioned Medium

We have examined the in vitro interaction between bronchial epithelial cells and eosinophils derived from five asthmatics by determining the effect of epithelial cell-conditioned medium on the survival and activation of peripheral blood eosinophils. The supernatants of epithelial cells from six normal donors were used as control. The asthmatic epithelial cell-conditioned medium significantly increased the survival of eosinophils cultured for 3 (P less than 0.025) and 6 (P less than 0.05) days. The incubation of eosinophils with the supernatants of asthmatic epithelial cells for 1 h also increased the generation of superoxide anion and the release of leukotriene C4, triggered by phorbol myristate acetate and calcium ionophore, by more than twofold. The preincubation of asthmatic epithelial cell-conditioned media with saturating concentrations of a mono-specific antiserum against granulocyte-macrophage colony-stimulating factor (GM-CSF) completely abolished their activity, whereas the addition of recombinant human GM-CSF restored it. The supernatants of asthmatic epithelial cells contained 0.88 +/- 0.09 (SD) ng/5 X 10(5) cells immunoreactive GM-CSF, and this amount was significantly greater than that measured in the supernatants of normal epithelial cells (0.21 +/- 0.105, P less than 0.025). Bronchial epithelial cells from asthmatics also expressed increased levels of GM-CSF mRNA when compared with normal epithelial cells. Thus both the synthesis and release of GM-CSF by bronchial epithelial cells are upregulated in asthma, and this may contribute to the persistence of eosinophil infiltration and activation in asthmatic airways.

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