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1.
Nat Commun ; 12(1): 1536, 2021 03 09.
Artículo en Inglés | MEDLINE | ID: mdl-33750776

RESUMEN

Hyperactivation of the MAPK signaling pathway motivates the clinical use of MAPK inhibitors for BRAF-mutant melanomas. Heterogeneity in differentiation state due to epigenetic plasticity, however, results in cell-to-cell variability in the state of MAPK dependency, diminishing the efficacy of MAPK inhibitors. To identify key regulators of such variability, we screen 276 epigenetic-modifying compounds, individually or combined with MAPK inhibitors, across genetically diverse and isogenic populations of melanoma cells. Following single-cell analysis and multivariate modeling, we identify three classes of epigenetic inhibitors that target distinct epigenetic states associated with either one of the lysine-specific histone demethylases Kdm1a or Kdm4b, or BET bromodomain proteins. While melanocytes remain insensitive, the anti-tumor efficacy of each inhibitor is predicted based on melanoma cells' differentiation state and MAPK activity. Our systems pharmacology approach highlights a path toward identifying actionable epigenetic factors that extend the BRAF oncogene addiction paradigm on the basis of tumor cell differentiation state.


Asunto(s)
Diferenciación Celular/efectos de los fármacos , Epigenómica/métodos , Melanoma/metabolismo , Dependencia del Oncogén , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Epigénesis Genética/efectos de los fármacos , Femenino , Histona Demetilasas/metabolismo , Humanos , Histona Demetilasas con Dominio de Jumonji/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Melanocitos/metabolismo , Melanoma/genética , Ratones , Ratones Desnudos , Mutación , Dependencia del Oncogén/genética , Inhibidores de Proteínas Quinasas/farmacología , Proteínas Proto-Oncogénicas B-raf/genética , Transducción de Señal/efectos de los fármacos , Ensayos Antitumor por Modelo de Xenoinjerto
2.
Ecotoxicol Environ Saf ; 214: 112080, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33677380

RESUMEN

Resveratrol (RES) is a natural polyphenolic compound with a broad range of physiological and pharmacological properties. Previous studies have shown that RES also plays an important role in protecting and promoting early bone metabolism and differentiation. The accumulation of cadmium (Cd), one of the world's most poisonous substances, can inhibit skeletal growth and bone maturation, thus causing osteoporosis. However, whether RES can prevent the Cd-induced inhibition of osteogenic differentiation remains unknown. In this study, we found that RES promoted the early maturity of osteoblastic MC3T3-E1 cells, as demonstrated by the significantly increased mRNA and protein expression of a range of differentiation markers, including alkaline phosphatase (ALP), collagen 1 (COL1), bone morphogenetic protein-2 (BMP-2), and runt-related transcription factor 2 (RUNX2). In contrast, we found that cadmium chloride (CdCl2) inhibited the viability and osteogenic maturity of MC3T3-E1 cells. We also demonstrated that RES pretreatment for 30 min provided significant protection against Cd-induced apoptosis and attenuated the inhibition of osteogenic differentiation induced by Cd by modulating ERK1/2 and JNK signaling. In conclusion, our results indicate that RES is a potential femoral protectant that not only enhance the viability and early differentiation of osteoblasts, but also protect osteoblasts from cadmium damage.


Asunto(s)
Cadmio/toxicidad , Sustancias Protectoras/farmacología , Resveratrol/farmacología , Fosfatasa Alcalina/metabolismo , Animales , Proteína Morfogenética Ósea 2 , Cadmio/metabolismo , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal , Sistema de Señalización de MAP Quinasas , Proteína Quinasa 3 Activada por Mitógenos , Osteoblastos/citología , Osteogénesis/genética
3.
Molecules ; 26(4)2021 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-33669807

RESUMEN

Dental papilla cells (DPCs), precursors of odontoblasts, are considered promising seed cells for tissue engineering. Emerging evidence suggests that melatonin promotes odontoblastic differentiation of DPCs and affects tooth development, although the precise mechanisms remain unknown. Retinoid acid receptor-related orphan receptor α (RORα) is a nuclear receptor for melatonin that plays a critical role in cell differentiation and embryonic development. This study aimed to explore the role of RORα in odontoblastic differentiation and determine whether melatonin exerts its pro-odontogenic effect via RORα. Herein, we observed that RORα was expressed in DPCs and was significantly increased during odontoblastic differentiation in vitro and in vivo. The overexpression of RORα upregulated the expression of odontogenic markers, alkaline phosphatase (ALP) activity and mineralized nodules formation (p < 0.05). In contrast, odontoblastic differentiation of DPCs was suppressed by RORα knockdown. Moreover, we found that melatonin elevated the expression of odontogenic markers, which was accompanied by the upregulation of RORα (p < 0.001). Utilising small interfering RNA, we further demonstrated that RORα inhibition attenuated melatonin-induced odontogenic gene expression, ALP activity and matrix mineralisation (p < 0.01). Collectively, these results provide the first evidence that RORα can promote odontoblastic differentiation of DPCs and mediate the pro-odontogenic effect of melatonin.


Asunto(s)
Diferenciación Celular , Papila Dental/citología , Melatonina/farmacología , Miembro 1 del Grupo F de la Subfamilia 1 de Receptores Nucleares/metabolismo , Odontoblastos/citología , Odontoblastos/metabolismo , Odontogénesis , Animales , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Odontoblastos/efectos de los fármacos , Odontogénesis/efectos de los fármacos , Ratas Sprague-Dawley , Regulación hacia Arriba/efectos de los fármacos
4.
Int J Nanomedicine ; 16: 1509-1523, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33658781

RESUMEN

Purpose: The study was intended to create a uniform zirconia layer even on the surface of complex structures via atomic layer deposition (ALD). The impact of crystalline zirconia deposited by ALD on bacterial adhesion and osteoblast viability was assessed via surface treatment of dental implants. Methods: Amorphous zirconia was deposited using an atomic layer deposition reactor (Atomic Classic, CN1, Hwaseong, Korea) on titanium discs. Heating the samples at 400°C resulted in crystallization. Samples were divided into three groups: the control group, the group carrying amorphous ALD-zirconia (Z group), and the heat-treated group following zirconia ALD deposition (ZH group).The surface of each sample was analyzed, followed by the assessment of adhesion of Streptococcus mutans and Porphyromonas gingivalis, and viability and differentiation of MC3T3-E1 cells. Results: The adhesion of S. mutans and P. gingivalis was significantly reduced in the Z and ZH groups compared with the control group (P < 0.05). The viability of MC3T3-E1 cells was significantly increased in the ZH group compared with the control group (P < 0.001), while no significant differences were observed in the Z group (P > 0.05). Differentiation of MC3T3-E1 cells showed a marginally significant increase in the ZH group compared with the control group (P < 0.1), while no significant differences were found in the Z group (P > 0.1). Conclusion: Compared with the pure titanium group, the groups that were coated with zirconia via ALD showed a decreased adhesion of S. mutans during the early stages of biofilm formation and P. gingivalis adhesion inducing peri-implantitis, and an increase in MC3T3-E1 cell viability and differentiation. The findings indicate the possibility of treating the implant surface to reduce peri-implantitis and improve osseointegration.


Asunto(s)
Adhesión Bacteriana , Osteoblastos/citología , Titanio/farmacología , Circonio/química , Animales , Adhesión Bacteriana/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Humanos , Ratones , Microscopía de Fuerza Atómica , Osteoblastos/efectos de los fármacos , Osteoblastos/metabolismo , Osteoblastos/ultraestructura , Espectroscopía de Fotoelectrones , Espectrometría por Rayos X , Propiedades de Superficie , Difracción de Rayos X
5.
Ecotoxicol Environ Saf ; 214: 112121, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33721578

RESUMEN

Perfluorooctane sulfonate is related to male reproductive dysfunction in rats and humans. However, the underlying mechanism remains unknown. Here, we reported the effects of short-term exposure to perfluorooctane sulfonate on the regeneration of Leydig cells in vivo and investigated possible mechanisms in vitro. After adult male Sprague-Dawley rats were gavaged perfluorooctane sulfonate (0, 5 or 10 mg/kg/day) for 7 days and then injected intraperitoneally ethane dimethane sulfonate next day to eliminate Leydig cells, the Leydig cell regeneration process was monitored. Perfluorooctane sulfonate significantly lowered serum testosterone levels, reduced the number of regenerated Leydig cells, down-regulated the expression of Lhcgr, Scarb1, Star, Cyp11a1, Hsd3b1, Cyp17a1, and Dhh) and their proteins at doses of 5 and 10 mg/kg 35 and 56 days after ethane dimethane sulfonate. Using a 3D seminiferous tubule culture system to study the development of stem Leydig cells, we found that perfluorooctane sulfonate inhibited stem Leydig cell proliferation and differentiation and hedgehog signaling pathway. In conclusion, a short-term exposure to perfluorooctane sulfonate can inhibit the development of stem Leydig cells into the Leydig cell lineage via direct suppression of hedgehog signaling pathway and indirect inhibition of desert hedgehog section by Sertoli cells.


Asunto(s)
Ácidos Alcanesulfónicos/toxicidad , Fluorocarburos/toxicidad , Testículo/efectos de los fármacos , Animales , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Proteínas Hedgehog/metabolismo , Masculino , Mesilatos , Ratas Sprague-Dawley , Regeneración , Transducción de Señal/efectos de los fármacos , Testículo/citología , Testículo/metabolismo , Testículo/fisiología , Testosterona/sangre
6.
Carbohydr Polym ; 260: 117769, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712127

RESUMEN

Periodontal defect poses a significant challenge in orthopedics. Guided Bone Regeneration (GBR) membrane is considered as one of the most successful methods applied to reconstruct alveolar bone and then to achieve periodontal defect repair/regeneration. In this paper, a novel polyamide-6/chitosan@nano-hydroxyapatite/polyamide-6 (PA6/CS@n-HA/PA6) bilayered tissue guided membranes by combining a solvent casting and an electrospinning technique was designed. The developed PA6/CS@n-HA/PA6 composites were characterized by a series of tests. The results show that n-HA/PA6 and electrospun PA6/CS layers are tightly bound by molecular interaction and chemical bonding, which enhances the bonding strength between two distinct layers. The porosity and adsorption average pore diameter of the PA6/CS@n-HA/PA6 membranes are 36.90 % and 22.61 nm, respectively. The tensile strength and elastic modulus of PA6/CS@n-HA/PA6 composites are 1.41 ± 0.18 MPa and 7.15 ± 1.09 MPa, respectively. In vitro cell culture studies demonstrate that PA6/CS@n-HA/PA6 bilayered scaffolds have biological safety, good bioactivity, biocompatibility and osteoconductivity.


Asunto(s)
Regeneración Ósea , Caprolactama/análogos & derivados , Quitosano/química , Durapatita/química , Membranas Artificiales , Nanoestructuras/química , Polímeros/química , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Regeneración Ósea/efectos de los fármacos , Caprolactama/química , Adhesión Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Ratones , Nanofibras/química , Nanoestructuras/toxicidad , Porosidad , Propiedades de Superficie , Resistencia a la Tracción
7.
Carbohydr Polym ; 260: 117780, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712136

RESUMEN

In this study, we prepared a biomimetic hyaluronic acid oligosaccharides (oHAs)-based composite scaffold to develop a bone tissue-engineered scaffold for stimulating osteogenesis and endothelialization. The functional oHAs products were firstly synthesized, namely collagen/hyaluronic acid oligosaccharides/hydroxyapatite (Col/oHAs/HAP), chitosan/hyaluronic acid oligosaccharides (CTS/oHAs), and then uniformly distributed in poly (lactic-co-glycolic acid) (PLGA) solution followed by freeze-drying to obtain three-dimensional interconnected scaffolds as temporary templates for bone regeneration. The morphology, physicochemical properties, compressive strength, and degradation behavior of the fabricated scaffolds, as well as in vitro cell responses seeded on these scaffolds and in vivo biocompatibility, were investigated to evaluate the potential for bone tissue engineering. The results indicated that the oHAs-based scaffolds can promote the attachment of endothelial cells, facilitate the osteogenic differentiation of MC3T3-E1 and BMSCs, and have ideal biocompatibility and tissue regenerative capacity, suggesting their potential to serve as alternative candidates for bone tissue engineering applications.


Asunto(s)
Materiales Biocompatibles/química , Quitosano/química , Colágeno/química , Ingeniería de Tejidos , Animales , Materiales Biocompatibles/farmacología , Diferenciación Celular/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Durapatita/química , Ácido Hialurónico/química , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Ratones , Oligosacáridos/química , Osteoblastos/citología , Osteoblastos/metabolismo , Osteogénesis/efectos de los fármacos , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Andamios del Tejido/química
8.
Cell Prolif ; 54(4): e13012, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33656760

RESUMEN

OBJECTIVES: Vitronectin (VTN) has been widely used for the maintenance and expansion of human pluripotent stem cells (hPSCs) as feeder-free conditions. However, the effect of VTN on hPSC differentiation remains unclear. Here, we investigated the role of VTN in early haematopoietic development of hPSCs. MATERIALS AND METHODS: A chemically defined monolayer system was applied to study the role of different matrix or basement membrane proteins in haematopoietic development of hPSCs. The role of integrin signalling in VTN-mediated haematopoietic differentiation was investigated by integrin antagonists. Finally, small interfering RNA was used to knock down integrin gene expression in differentiated cells. RESULTS: We found that the haematopoietic differentiation of hPSCs on VTN was far more efficient than that on Matrigel that is also often used for hPSC culture. VTN promoted the fate determination of endothelial-haematopoietic lineage during mesoderm development to generate haemogenic endothelium (HE). Moreover, we demonstrated that the signals through αvß3 and αvß5 integrins were required for VTN-promoted haematopoietic differentiation. Blocking αvß3 and αvß5 integrins by the integrin antagonists impaired the development of HE, but not endothelial-to-haematopoietic transition (EHT). Finally, both αvß3 and αvß5 were confirmed acting synergistically for early haematopoietic differentiation by knockdown the expression of αv, ß3 or ß5. CONCLUSION: The established VTN-based monolayer system of haematopoietic differentiation of hPSCs presents a valuable platform for further investigating niche signals involved in human haematopoietic development.


Asunto(s)
Diferenciación Celular/efectos de los fármacos , Integrina alfaVbeta3/metabolismo , Receptores de Vitronectina/metabolismo , Vitronectina/farmacología , Adhesión Celular/efectos de los fármacos , Línea Celular , Regulación de la Expresión Génica , Células Madre Hematopoyéticas/citología , Células Madre Hematopoyéticas/metabolismo , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Humanos , Integrina alfaVbeta3/antagonistas & inhibidores , Integrina alfaVbeta3/genética , Mesodermo/citología , Mesodermo/crecimiento & desarrollo , Mesodermo/metabolismo , Células Madre Pluripotentes/citología , Células Madre Pluripotentes/metabolismo , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Receptores de Vitronectina/antagonistas & inhibidores , Receptores de Vitronectina/genética , Transducción de Señal/efectos de los fármacos , Venenos de Serpiente/farmacología
9.
Methods Mol Biol ; 2265: 119-128, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33704710

RESUMEN

Tumor-associated macrophages (TAMs) are one of most important components of the tumor microenvironment. Although many assays have been developed to differentiate monocytes into macrophages (Mϕ) for studying the biology of TAMs in vitro, little is known whether the macrophages induced by these approaches can recapitulate the biology of TAMs present in the tumor microenvironment. We have developed a novel assay to differentiate human monocytes into TAMs using modified melanoma-conditioned medium, which is derived from the concentrated tumor cell culture medium. Characterization of these modified melanoma-conditioned medium-induced macrophages (MCMI-Mϕ) by multiple flow cytometry, Luminex, microarray, and immunohistochemistry analyses indicates that MCMI-Mϕ are phenotypically and functionally highly similar to the TAMs present in the tumor microenvironment.


Asunto(s)
Diferenciación Celular/efectos de los fármacos , Medios de Cultivo Condicionados/farmacología , Macrófagos/citología , Melanoma/metabolismo , Técnicas de Cultivo de Célula/métodos , Línea Celular Tumoral , Humanos , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/efectos de los fármacos , Melanoma/patología , Monocitos/citología , Monocitos/efectos de los fármacos , Microambiente Tumoral
10.
Methods Mol Biol ; 2273: 151-158, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33604851

RESUMEN

The first differentiation event in mammalian embryos is the formation of the trophectoderm, which is the progenitor of the outer epithelial component of the placenta and supports the fetus during intrauterine life. Our understanding of these events is limited, particularly in human, because of ethical and legal restrictions and availability of adequate in vitro models would be very advantageous. Here we describe a method that converts human fibroblasts into trophoblast-like cells, combining the use of 5-azacytidine-CR (5-aza-CR) to erase the original cell phenotype and a cocktail containing bone morphogenetic protein 4 (BMP4) with inhibitors of the Activin/Nodal/ERK signaling pathways, to drive erased fibroblasts into the trophoblastic differentiation. This innovative method uses very easily accessible cells to derive trophoblast-like cells and it can be useful to study embryo implantation disorders related to aging.


Asunto(s)
Técnicas de Cultivo de Célula/métodos , Fibroblastos/citología , Trofoblastos/citología , Activinas/antagonistas & inhibidores , Animales , Azacitidina/farmacología , Proteína Morfogenética Ósea 4/metabolismo , Proteína Morfogenética Ósea 4/farmacología , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/genética , Implantación del Embrión , Embrión de Mamíferos/metabolismo , Células Madre Embrionarias/citología , Femenino , Fibroblastos/efectos de los fármacos , Fibroblastos/fisiología , Humanos , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Proteína Nodal/antagonistas & inhibidores , Placenta/citología , Embarazo , Transducción de Señal , Piel/citología , Piel/crecimiento & desarrollo
11.
Life Sci ; 272: 119157, 2021 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-33524418

RESUMEN

Stem cell-based therapy is known as a regenerative approach for a variety of diseases and tissue injuries. These cells exert their therapeutic effects through paracrine secretions namely extracellular vesicles. To achieve higher therapeutic potential, a variety of delivery routes have been tested in clinical and preclinical studies. Direct cell injection, intra-venous administration, and intra-arterial infusion are widely used methods of stem cells delivery but these methods are associated with several complications. As one of the most popular biological delivery systems, amniotic membrane has been widely utilized to support cell proliferation and differentiation therefore facilitating tissue regeneration without endangering the stem cells' viability. It is composed of several extracellular matrix components and growth factors. Due to these characteristics, amniotic membrane can mimic the stem cell's niche and can be an ideal carrier for stem cell transplantation. Here, we provide an overview of the recent progress, challenges, and future perspectives in the use of amniotic membrane as a delivery platform for stem cells.


Asunto(s)
Amnios/metabolismo , Sistemas de Liberación de Medicamentos/métodos , Trasplante de Células Madre/métodos , Amnios/fisiología , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Proliferación Celular/efectos de los fármacos , Proliferación Celular/fisiología , Vesículas Extracelulares/metabolismo , Humanos , Células Madre Mesenquimatosas/citología , Medicina Regenerativa/métodos , Células Madre/citología
12.
Methods Mol Biol ; 2235: 47-59, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33576970

RESUMEN

We report the use of self-assembled peptide (F2/S) hydrogels and cellular metabolomics to identify a number of innate molecules that are integral to the metabolic processes which drive cellular differentiation of multipotent pericyte stem cells. The culture system relies solely on substrate mechanics to induce differentiation in the absence of traditional differentiation media and therefore is a non-invasive approach to assessing cellular behavior at the molecular level and identifying key metabolites in this process. This novel approach demonstrates that simple metabolites can provide an alternative means to direct stem cell differentiation and that biomaterials can be used to identify them simply and quickly.


Asunto(s)
Metabolómica/métodos , Pericitos/citología , Pericitos/trasplante , Animales , Materiales Biocompatibles/metabolismo , Capilares/citología , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Células Cultivadas , Células Endoteliales/citología , Humanos , Hidrogeles/química , Microvasos/citología , Células Madre Multipotentes/efectos de los fármacos , Péptidos/química , Pericitos/metabolismo , Fenotipo
13.
Methods Mol Biol ; 2235: 119-125, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33576973

RESUMEN

Human pluripotent stem cells (hPSCs), either embryonic or induced, offer a plentiful platform for derivation of multiple cell types. Pericytes, generated from hPSCs, are multipotent precursors with vasculogenic features that exhibit high proliferation capability in long-term cultures. Administration of hPSC-pericytes into ischemic murine hind limb is associated with therapeutic angiogenesis and attenuation of muscle wasting. Here, we describe the protocol for derivation of large numbers of pericytes from spontaneously differentiating hPSC-embryoid bodies.


Asunto(s)
Técnicas de Cultivo de Célula/métodos , Diferenciación Celular/fisiología , Pericitos/metabolismo , Animales , Diferenciación Celular/efectos de los fármacos , Línea Celular , Humanos , Pericitos/citología , Células Madre Pluripotentes/citología , Células Madre Pluripotentes/metabolismo
14.
Methods Mol Biol ; 2235: 127-137, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33576974

RESUMEN

Human pericytes are a perivascular cell population with mesenchymal stem cell properties, present in all vascularized tissues. Human pericytes have a distinct immunoprofile, which may be leveraged for purposes of cell purification. Adipose tissue is the most commonly used cell source for human pericyte derivation. Pericytes can be isolated by FACS (fluorescence-activated cell sorting), most commonly procured from liposuction aspirates. Pericytes have clonal multilineage differentiation potential, and their potential utility for bone regeneration has been described across multiple animal models. The following review will discuss in vivo methods for assessing the bone-forming potential of purified pericytes. Potential models include (1) mouse intramuscular implantation, (2) mouse calvarial defect implantation, and (3) rat spinal fusion models. In addition, the presented surgical protocols may be used for the in vivo analysis of other osteoprogenitor cell types.


Asunto(s)
Células de la Médula Ósea/metabolismo , Pericitos/metabolismo , Ingeniería de Tejidos/métodos , Tejido Adiposo/citología , Animales , Células de la Médula Ósea/citología , Regeneración Ósea/fisiología , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Línea Celular , Separación Celular/métodos , Humanos , Células Madre Mesenquimatosas/citología , Ratones , Osteogénesis/fisiología , Pericitos/citología , Ratas
15.
Molecules ; 26(3)2021 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-33573260

RESUMEN

Phytoestrogens are herbal polyphenolic compounds that exert various estrogen-like effects in animals and can be taken in easily from a foodstuff in daily life. The fallopian tube lumen, where transportation of the oocyte occurs, is lined with secretory cells and multi-ciliated epithelial cells. Recently, we showed that estrogen induces multi-ciliogenesis in the porcine fallopian tube epithelial cells (FTECs) through the activation of the estrogen receptor beta (ERß) pathway and simultaneous inhibition of the Notch pathway. Thus, ingested phytoestrogens may induce FTEC ciliogenesis and thereby affect the fecundity. To address this issue, we added isoflavones (genistein, daidzein, or glycitin) and coumestan (coumestrol) to primary culture FTECs under air-liquid interface conditions and assessed the effects of each compound. All phytoestrogens except glycitin induced multi-ciliated cell differentiation, which followed Notch signal downregulation. On the contrary, the differentiation of secretory cells decreased slightly. Furthermore, genistein and daidzein had a slight effect on the proportion of proliferating cells exhibited by Ki67 expression. Ciliated-cell differentiation is inhibited by the ERß antagonist, PHTPP. Thus, this study suggests that phytoestrogens can improve the fallopian tube epithelial sheet homeostasis by facilitating the genesis of multi-ciliated cells and this effect depends on the ERß-mediated pathway.


Asunto(s)
Epitelio/crecimiento & desarrollo , Receptor beta de Estrógeno/genética , Fitoestrógenos/farmacología , Polifenoles/farmacología , Animales , Biomimética , Diferenciación Celular/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Epitelio/efectos de los fármacos , Trompas Uterinas/efectos de los fármacos , Trompas Uterinas/crecimiento & desarrollo , Femenino , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Humanos , Porcinos
16.
Nat Commun ; 12(1): 1248, 2021 02 23.
Artículo en Inglés | MEDLINE | ID: mdl-33623001

RESUMEN

Mutations in human equilibrative nucleoside transporter 3 (ENT3) encoded by SLC29A3 results in anemia and erythroid hypoplasia, suggesting that ENT3 may regulate erythropoiesis. Here, we demonstrate that lysosomal ENT3 transport of taurine-conjugated bile acids (TBA) facilitates TBA chemical chaperone function and alleviates endoplasmic reticulum (ER) stress in expanding mouse hematopoietic stem and progenitor cells (HSPCs). Slc29a3-/- HSPCs accumulate less TBA despite elevated levels of TBA in Slc29a3-/- mouse plasma and have elevated basal ER stress, reactive oxygen species (ROS), and radiation-induced apoptosis. Reintroduction of ENT3 allows for increased accumulation of TBA into HSPCs, which results in TBA-mediated alleviation of ER stress and erythroid apoptosis. Transplanting TBA-preconditioned HSPCs expressing ENT3 into Slc29a3-/- mice increase bone marrow repopulation capacity and erythroid pool size and prevent early mortalities. Together, these findings suggest a putative role for a facilitative lysosomal transporter in the bile acid regulation of ER stress in mouse HSPCs which may have implications in erythroid biology, the treatment of anemia observed in ENT3-mutated human genetic disorders, and nucleoside analog drug therapy.


Asunto(s)
Ácidos y Sales Biliares/metabolismo , Estrés del Retículo Endoplásmico , Células Madre Hematopoyéticas/metabolismo , Lisosomas/metabolismo , Animales , Apoptosis/efectos de los fármacos , Ácidos y Sales Biliares/sangre , Transporte Biológico/efectos de los fármacos , Células de la Médula Ósea/metabolismo , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Células Eritroides/efectos de los fármacos , Células Eritroides/metabolismo , Trasplante de Células Madre Hematopoyéticas , Concentración de Iones de Hidrógeno , Lisosomas/efectos de los fármacos , Metabolómica , Ratones , Proteínas de Transporte de Nucleósidos/metabolismo , Taurina/metabolismo , Ácido Tauroquenodesoxicólico/farmacología
17.
J Vis Exp ; (167)2021 01 22.
Artículo en Inglés | MEDLINE | ID: mdl-33554962

RESUMEN

Colorectal cancers are characterized by heterogeneity and a hierarchical organization comprising a population of cancer stem cells (CSCs) responsible for tumor development, maintenance, and resistance to drugs. A better understanding of CSC properties for their specific targeting is, therefore, a pre-requisite for effective therapy. However, there is a paucity of suitable preclinical models for in-depth investigations. Although in vitro two-dimensional (2D) cancer cell lines provide valuable insights into tumor biology, they do not replicate the phenotypic and genetic tumor heterogeneity. In contrast, three-dimensional (3D) models address and reproduce near-physiological cancer complexity and cell heterogeneity. The aim of this work was to design a robust and reproducible 3D culture system to study CSC biology. The present methodology describes the development and optimization of conditions to generate 3D spheroids, which are homogenous in size, from Caco2 colon adenocarcinoma cells, a model that can be used for long-term culture. Importantly, within the spheroids, the cells which were organized around lumen-like structures, were characterized by differential cell proliferation patterns and by the presence of CSCs expressing a panel of markers. These results provide the first proof-of-concept for the appropriateness of this 3D approach to study cell heterogeneity and CSC biology, including the response to chemotherapy.


Asunto(s)
Neoplasias del Colon/patología , Células Madre Neoplásicas/patología , Esferoides Celulares/patología , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Biomarcadores de Tumor/metabolismo , Células CACO-2 , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Neoplasias del Colon/tratamiento farmacológico , Enterocitos/efectos de los fármacos , Enterocitos/patología , Técnica del Anticuerpo Fluorescente , Humanos , Células Madre Neoplásicas/efectos de los fármacos , Esferoides Celulares/efectos de los fármacos , Coloración y Etiquetado
18.
Nat Commun ; 12(1): 1031, 2021 02 15.
Artículo en Inglés | MEDLINE | ID: mdl-33589620

RESUMEN

The application of physical stimuli to cell cultures has shown potential to modulate multiple cellular functions including migration, differentiation and survival. However, the relevance of these in vitro models to future potential extrapolation in vivo depends on whether stimuli can be applied "externally", without invasive procedures. Here, we report on the fabrication and exploitation of dynamic additive-manufactured Janus scaffolds that are activated on-command via external application of ultrasounds, resulting in a mechanical nanovibration that is transmitted to the surrounding cells. Janus scaffolds were spontaneously formed via phase-segregation of biodegradable polycaprolactone (PCL) and polylactide (PLA) blends during the manufacturing process and behave as ultrasound transducers (acoustic to mechanical) where the PLA and PCL phases represent the active and backing materials, respectively. Remote stimulation of Janus scaffolds led to enhanced cell proliferation, matrix deposition and osteogenic differentiation of seeded human bone marrow derived stromal cells (hBMSCs) via formation and activation of voltage-gated calcium ion channels.


Asunto(s)
Plásticos Biodegradables/farmacología , Mecanotransducción Celular , Células Madre Mesenquimatosas/efectos de los fármacos , Poliésteres/farmacología , Andamios del Tejido , Plásticos Biodegradables/química , Regeneración Ósea/genética , Huesos/citología , Huesos/metabolismo , Canales de Calcio Activados por la Liberación de Calcio/fisiología , Diferenciación Celular/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Humanos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Poliésteres/química , Impresión Tridimensional , Ingeniería de Tejidos/métodos , Ondas Ultrasónicas
19.
Yonsei Med J ; 62(2): 137-148, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33527793

RESUMEN

PURPOSE: In organ transplantation, the need for immune modulation rather than immune suppression has been emphasized. In this study, we investigated whether combinatorial treatments of with thalidomide (TM) and dexamethasone (DX) might be new approaches to induce systemic immunomodulation on T cells and other immune cells that regulate the expression of co-inhibitory molecules. MATERIALS AND METHODS: Naïve splenic T cells from C57BL/6 mice were sort-purified and cultured in vitro for CD4+ T cell proliferation and regulatory T cell (Treg) conversion in the presence of TM or/and DX. Expression of cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) and programmed death-1 (PD-1) in proliferated and converted T cells was quantified by flow cytometry. We also quantified in vivo expression of CTLA-4 and PD-1 on splenic CD4+ T cells and other immune cells isolated from TM- or/and DX-treated mice. Mixed lymphocytes reactions (MLR) were performed to evaluate the capacity of immune cells in carrying out immune responses. RESULTS: CTLA-4 expressions in effector T cells in vivo and in Tregs in vivo/vitro significantly increased upon TM/DX combinatorial treatment. Corresponding to increased CTLA-4 expression in T cells, the expression of ligand molecules for CTLA-4 significantly increased in splenic dendritic cells in TM/DX-treated groups. In addition, MLR results demonstrated that splenocytes isolated from TM/DX-treated mice significantly suppressed the proliferation of T cells isolated from other strains. CONCLUSION: Based on these results, we suggest that TM/DX combinatorial treatments might be efficient immunomodulatory methods for regulating T cell immunity.


Asunto(s)
Dexametasona/farmacología , Inmunomodulación/efectos de los fármacos , Linfocitos T/inmunología , Talidomida/farmacología , Animales , Antígeno CTLA-4/metabolismo , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Dendríticas/efectos de los fármacos , Células Dendríticas/metabolismo , Citometría de Flujo , Activación de Linfocitos/efectos de los fármacos , Prueba de Cultivo Mixto de Linfocitos , Masculino , Ratones Endogámicos C57BL , Receptor de Muerte Celular Programada 1/metabolismo , Bazo/citología , Linfocitos T/efectos de los fármacos , Linfocitos T Reguladores/inmunología
20.
Nat Commun ; 12(1): 723, 2021 02 01.
Artículo en Inglés | MEDLINE | ID: mdl-33526787

RESUMEN

Bone metastatic prostate cancer (PCa) promotes mesenchymal stem cell (MSC) recruitment and their differentiation into osteoblasts. However, the effects of bone-marrow derived MSCs on PCa cells are less explored. Here, we report MSC-derived interleukin-28 (IL-28) triggers prostate cancer cell apoptosis via IL-28 receptor alpha (IL-28Rα)-STAT1 signaling. However, chronic exposure to MSCs drives the selection of prostate cancer cells that are resistant to IL-28-induced apoptosis and therapeutics such as docetaxel. Further, MSC-selected/IL-28-resistant prostate cancer cells grow at accelerated rates in bone. Acquired resistance to apoptosis is PCa cell intrinsic, and is associated with a shift in IL-28Rα signaling via STAT1 to STAT3. Notably, STAT3 ablation or inhibition impairs MSC-selected prostate cancer cell growth and survival. Thus, bone marrow MSCs drive the emergence of therapy-resistant bone metastatic prostate cancer yet this can be disabled by targeting STAT3.


Asunto(s)
Adenocarcinoma/secundario , Neoplasias Óseas/secundario , Células Madre Mesenquimatosas/patología , Neoplasias de la Próstata/patología , Receptores de Interferón/metabolismo , Ácidos Aminosalicílicos/farmacología , Ácidos Aminosalicílicos/uso terapéutico , Animales , Apoptosis/efectos de los fármacos , Bencenosulfonatos/farmacología , Bencenosulfonatos/uso terapéutico , Neoplasias Óseas/tratamiento farmacológico , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular , Medios de Cultivo Condicionados/metabolismo , Modelos Animales de Enfermedad , Docetaxel/farmacología , Docetaxel/uso terapéutico , Humanos , Interferones/genética , Interferones/metabolismo , Masculino , Ratones Noqueados , Osteoblastos/patología , Cultivo Primario de Células , Neoplasias de la Próstata/tratamiento farmacológico , ARN Interferente Pequeño/metabolismo , Receptores de Interferón/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Factor de Transcripción STAT1/metabolismo , Factor de Transcripción STAT3/antagonistas & inhibidores , Factor de Transcripción STAT3/metabolismo , Tibia/patología
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