Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 20.935
Filtrar
1.
Anticancer Res ; 41(9): 4259-4269, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34475045

RESUMO

BACKGROUND: Compared to two-dimensional cultures, three-dimensional (3D) cultures have many advantages in cancer studies. Nevertheless, their implementation is unsatisfactory. This study aimed to develop an anchorage-dependent 3D culture model for colorectal cancer research. MATERIALS AND METHODS: Human HCT116, DLD-1 and SW620 colorectal cell lines were cultured in a gelatin sponge, and its applicability for morphological examination was studied. RESULTS: The resulting specimens were suitable for scanning electron microscopy, transmission electron microscopy, and immunohistochemical examination. HCT116 formed smaller structures and migrated through the pores of the sponge. DLD-1 formed larger structures with tight cell-to-cell adhesion. SW620 also formed large structures but small clustered cells tended to attach to the anchorage more favorably. Immunohistochemical staining demonstrated phosphorylated yes-associated protein (YAP) localized near the attachment site in HCT116 cells. CONCLUSION: Because the gelatin sponge provided suitable anchorage and the cultured cells formed distinguishable 3D structures, this method may be useful for further colorectal cancer research.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Técnicas de Cultura de Células/métodos , Neoplasias Colorretais/patologia , Gelatina/química , Tecidos Suporte/química , Fatores de Transcrição/metabolismo , Adesão Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Neoplasias Colorretais/metabolismo , Células HCT116 , Humanos , Fosforilação
2.
APMIS ; 129 Suppl 142: 1-30, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34399444

RESUMO

Over the past decade, 3D culture models of human and animal cells have found their way into tissue differentiation, drug development, personalized medicine and tumour behaviour studies. Embryoid bodies (EBs) are in vitro 3D cultures established from murine pluripotential stem cells, whereas tumoroids are patient-derived in vitro 3D cultures. This thesis aims to describe a new implication of an embryoid body model and to characterize the patient-specific microenvironment of the parental tumour in relation to tumoroid growth rate. In this thesis, we described a high-throughput monitoring method, where EBs are used as a dynamic angiogenesis model. In this model, digital image analysis (DIA) is implemented on immunohistochemistry (IHC) stained sections of the cultures over time. Furthermore, we have investigated the correlation between the genetic profile and inflammatory microenvironment of parental tumours on the in vitro growth rate of tumoroids. The EBs were cultured in spinner flasks. The samples were collected at days 4, 6, 9, 14, 18 and 21, dehydrated and embedded in paraffin. The histological sections were IHC stained for the endothelial marker CD31 and digitally scanned. The virtual whole-image slides were digitally analysed by Visiopharm® software. Histological evaluation showed vascular-like structures over time. The quantitative DIA was plausible to monitor significant increase in the total area of the EBs and an increase in endothelial differentiation. The tumoroids were established from 32 colorectal adenocarcinomas. The in vitro growth rate of the tumoroids was followed by automated microscopy over an 11-day period. The parental tumours were analysed by next-generation sequencing for KRAS, TP53, PIK3CA, SMAD4, MAP2K1, BRAF, FGFR3 and FBXW7 status. The tumoroids established from KRAS-mutated parental tumours showed a significantly higher growth rate compared to their wild-type counterparts. The density of CD3+ T lymphocytes and CD68+ macrophages was calculated in the centre of the tumours and at the invasive margin of the tumours. The high density of CD3+ cells and the low density of CD68+ cells showed a significant correlation with a higher growth rate of the tumoroids. In conclusion, a novel approach for histological monitoring of endothelial differentiation is presented in the stem cell-derived EBs. Furthermore, the KRAS status and density of CD3+ T cells and macrophages in the parental tumour influence the growth rate of the tumoroids. Our results indicate that these parameters should be included when tumoroids are to be implemented in personalized medicine.


Assuntos
Adenocarcinoma/patologia , Técnicas de Cultura de Células/métodos , Neoplasias Colorretais/patologia , Animais , Diferenciação Celular/fisiologia , Corpos Embrioides/patologia , Células Endoteliais/patologia , Humanos , Macrófagos/patologia , Camundongos , Células-Tronco/patologia , Linfócitos T/patologia , Microambiente Tumoral/fisiologia
3.
Nat Biomed Eng ; 5(8): 926-940, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34373601

RESUMO

Current protocols for the differentiation of human pluripotent stem cells (hPSCs) into chondrocytes do not allow for the expansion of intermediate progenitors so as to prospectively assess their chondrogenic potential. Here we report a protocol that leverages PRRX1-tdTomato reporter hPSCs for the selective induction of expandable and ontogenetically defined PRRX1+ limb-bud-like mesenchymal cells under defined xeno-free conditions, and the prospective assessment of the cells' chondrogenic potential via the cell-surface markers CD90, CD140B and CD82. The cells, which proliferated stably and exhibited the potential to undergo chondrogenic differentiation, formed hyaline cartilaginous-like tissue commensurate to their PRRX1-expression levels. Moreover, we show that limb-bud-like mesenchymal cells derived from patient-derived induced hPSCs can be used to identify therapeutic candidates for type II collagenopathy and we developed a method to generate uniformly sized hyaline cartilaginous-like particles by plating the cells on culture dishes coated with spots of a zwitterionic polymer. PRRX1+ limb-bud-like mesenchymal cells could facilitate the mass production of chondrocytes and cartilaginous tissues for applications in drug screening and tissue engineering.


Assuntos
Proteínas de Homeodomínio/genética , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Pluripotentes/citologia , Animais , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Condrócitos/citologia , Condrócitos/metabolismo , Condrócitos/transplante , Condrogênese , Doenças do Colágeno/terapia , Meios de Cultura/química , Proteínas de Homeodomínio/metabolismo , Humanos , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Células-Tronco Pluripotentes/metabolismo , Polímeros/química , Receptor beta de Fator de Crescimento Derivado de Plaquetas/metabolismo , Antígenos Thy-1/metabolismo , Engenharia Tecidual
6.
Int J Mol Sci ; 22(16)2021 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-34445681

RESUMO

Parthenogenetic embryos have been widely studied as an effective tool related to paternal and maternal imprinting genes and reproductive problems for a long time. In this study, we established a parthenogenetic epiblast-like stem cell line through culturing parthenogenetic diploid blastocysts in a chemically defined medium containing activin A and bFGF named paAFSCs. The paAFSCs expressed pluripotent marker genes and germ-layer-related genes, as well as being alkaline-phosphatase-positive, which is similar to epiblast stem cells (EpiSCs). We previously showed that advanced embryonic stem cells (ASCs) represent hypermethylated naive pluripotent embryonic stem cells (ESCs). Here, we converted paAFSCs to ASCs by replacing bFGF with bone morphogenetic protein 4 (BMP4), CHIR99021, and leukemia inhibitory factor (LIF) in a culture medium, and we obtained parthenogenetic advanced stem cells (paASCs). The paASCs showed similar morphology with ESCs and also displayed a stronger developmental potential than paAFSCs in vivo by producing chimaeras. Our study demonstrates that maternal genes could support parthenogenetic EpiSCs derived from blastocysts and also have the potential to convert primed state paAFSCs to naive state paASCs.


Assuntos
Células-Tronco Embrionárias/citologia , Células-Tronco Embrionárias/metabolismo , Partenogênese/fisiologia , Ativinas/metabolismo , Animais , Blastocisto/metabolismo , Proteína Morfogenética Óssea 4/farmacologia , Técnicas de Cultura de Células/métodos , Diferenciação Celular/efeitos dos fármacos , Metilação de DNA/efeitos dos fármacos , Técnicas de Cultura Embrionária/métodos , Feminino , Fatores de Crescimento de Fibroblastos/farmacologia , Camadas Germinativas/metabolismo , Camadas Germinativas/fisiologia , Fator Inibidor de Leucemia/farmacologia , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos ICR , Células-Tronco Embrionárias Murinas/citologia , Partenogênese/genética , Células-Tronco Pluripotentes/metabolismo , Células-Tronco Pluripotentes/patologia
7.
Int J Mol Sci ; 22(15)2021 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-34361088

RESUMO

Mouse embryonic stem cells (ESCs) are useful tools for studying early embryonic development and tissue formation in mammals. Since neural lineage differentiation is a major subject of organogenesis, the development of efficient techniques to induce neural stem cells (NSCs) from pluripotent stem cells must be preceded. However, the currently available NSC differentiation methods are complicated and time consuming. This study aimed to propose an efficient method for the derivation of NSCs from mouse ESCs; early neural lineage commitment was achieved using a three-dimensional (3D) culture system, followed by a two-dimensional (2D) NSC derivation. To select early neural lineage cell types during differentiation, Sox1-GFP transgenic ESCs were used. They were differentiated into early neural lineage, forming spherical aggregates, which were subsequently picked for the establishment of 2D NSCs. The latter showed a morphology similar to that of brain-derived NSCs and expressed NSC markers, Musashi, Nestin, N-cadherin, and Sox2. Moreover, the NSCs could differentiate into three subtypes of neural lineages, neurons, astrocytes, and oligodendrocytes. The results together suggested that ESCs could efficiently differentiate into tripotent NSCs through specification in 3D culture (for approximately 10 days) followed by 2D culture (for seven days).


Assuntos
Técnicas de Cultura de Células/métodos , Células-Tronco Embrionárias Murinas/citologia , Células-Tronco Neurais/citologia , Neurônios/citologia , Células-Tronco Pluripotentes/citologia , Animais , Diferenciação Celular , Células Cultivadas , Camundongos , Células-Tronco Embrionárias Murinas/metabolismo , Nestina/metabolismo , Células-Tronco Neurais/metabolismo , Neurônios/metabolismo , Células-Tronco Pluripotentes/metabolismo , Fatores de Transcrição SOXB1/metabolismo
8.
Biomolecules ; 11(7)2021 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-34208902

RESUMO

The vasculature of stem-cell-derived liver organoids can be engineered using methods that recapitulate embryonic liver development. Hepatic organoids with a vascular network offer great application prospects for drug screening, disease modeling, and therapeutics. However, the application of stem cell-derived organoids is hindered by insufficient vascularization and maturation. Here, we review different theories about the origin of hepatic cells and the morphogenesis of hepatic vessels to provide potential approaches for organoid generation. We also review the main protocols for generating vascularized liver organoids from stem cells and consider their potential and limitations in the generation of vascularized liver organoids.


Assuntos
Fígado/patologia , Organoides/irrigação sanguínea , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Avaliação Pré-Clínica de Medicamentos/métodos , Engenharia Genética/métodos , Hepatócitos/patologia , Humanos , Fígado/crescimento & desenvolvimento , Organogênese/fisiologia , Organoides/crescimento & desenvolvimento , Organoides/metabolismo , Células-Tronco/metabolismo
9.
Methods Mol Biol ; 2342: 737-763, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34272715

RESUMO

In the first edition of this book, we presented the basics of explicitly incorporating the lipid biochemistry into a confluent cell monolayer transport model and the novel findings of this model up to 2013, including the use of global optimization to fit the elementary rate constants and the efflux active P-glycoprotein (P-gp) membrane concentrations for the transport of four P-gp substrates across MDCKII-hMDR1-NKI confluent cell monolayers. This chapter is an update on that model, which has been focused primarily on discovering how microvilli morphology regulates the efflux active P-gp and the existence of, as yet, unidentified uptake transporters of P-gp substrates in all of the commonly used P-gp expressing cell lines used in the pharmaceutical industry, thereby adding new players to DDI predictions and IVIVE. The structural mass action kinetic model uses the general mass action reactions for P-gp binding and efflux, with the membrane structural parameters for the confluent cell monolayer to predict drug transport over time. Binding of drug to P-gp occurs within the cytosolic monolayer of the apical membrane, according to (a) the molar partition coefficient of the drug to the cytosolic monolayer and (b) the association rate constant, k1 (M-1 s-1), of the drug from the basolateral or apical outer monolayers into the P-gp binding site. Release of substrate from P-gp back into the cytosolic monolayer occurs with a dissociation rate constant kr (s-1) or, much less frequently, into the apical aqueous chamber with an efflux rate constant k2 (s-1). The model fits the efflux active P-gp concentration, T(0), i.e., the P-gp whose effluxed drug actually reaches the apical aqueous chamber, as opposed to the majority of P-gp whose effluxed drug is reabsorbed back into the same or neighboring microvilli prior to reaching the apical aqueous chamber. Efflux active P-gp largely resides near the tips of the microvilli. We have shown using kinetics and structured illumination microscopy that: (a) efflux active P-gp is controlled by microvilli morphology; (b) there are apical (AT) and basolateral (BT) uptake transporters for P-gp substrates in most, if not all, P-gp expressing cell lines used in the pharmaceutical industry, which exist, but which remain unidentified; (c) the lab-to-lab variability in P-gp IC50 values observed in the P-gp IC50 initiative was due to the conflated inhibition of P-gp and the basolateral digoxin uptake transporters by all 15 P-gp substrates tested in that study; (d) even the IC50 values for P-gp inhibition alone do not obey the Cheng-Prusoff relationship; (e) the fitted elementary rate constants and the molecular dissociation constant Ki for this kinetic model are system independent; and (f) the time dependence of product formation for these confluent cell monolayers is correlated with the P-gp Vmax/Km, when defined by its fitted elementary rate constants and uptake transporter clearances, without any steady-state assumptions.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/química , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Técnicas de Cultura de Células/métodos , Preparações Farmacêuticas/metabolismo , Animais , Sítios de Ligação , Transporte Biológico , Células CACO-2 , Células Cultivadas , Citosol/metabolismo , Humanos , Cinética , Microvilosidades/metabolismo , Modelos Teóricos
10.
Methods Mol Biol ; 2319: 51-60, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34331242

RESUMO

Cardiovascular disease is a worldwide health issue that affects millions of lives every year, and thus, researchers are in need of high-throughput model systems with which to investigate mechanisms of disease and to develop and test potential therapies. The use of human-derived induced pluripotent stem cells (iPSCs) differentiated into cardiomyocytes aims to address this need. While cardiac differentiation protocols have been established previously in iPSCs, optimization of cardiac differentiation remains crucial to obtaining high quality cardiomyocytes for future experimental analyses. Important factors to consider include cell density and rate of proliferation, temporal regulation of media changes throughout the differentiation process, and the concentration of the chemicals utilized. In this chapter, we present a detailed protocol to outline the process of differentiating cardiomyocytes from human iPSCs via modulation of Wnt signaling, characterization of cardiomyocytes by immunofluorescence, as well as guidelines for troubleshooting and optimizing these techniques.


Assuntos
Técnicas de Cultura de Células/métodos , Meios de Cultura/química , Células-Tronco Pluripotentes Induzidas/citologia , Desenvolvimento Muscular , Miócitos Cardíacos/citologia , Via de Sinalização Wnt , Imunofluorescência , Humanos , Técnicas In Vitro , Células-Tronco Pluripotentes Induzidas/metabolismo , Miócitos Cardíacos/metabolismo
11.
Methods Mol Biol ; 2319: 69-75, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34331244

RESUMO

There is increasing interest in the study of the mammalian lymphatic system, including the lymphatic endothelial cells (LECs) that make up lymphatic vessels. The ability to isolate primary LECs from tissue of normal and genetically modified mice permits detailed analysis of this unique cell type. Here, we describe a robust protocol for the isolation and in vitro expansion of LECs from mouse lung by antibody-based magnetic separation.


Assuntos
Técnicas de Cultura de Células/métodos , Separação Celular/métodos , Células Endoteliais/citologia , Separação Imunomagnética/métodos , Pulmão/citologia , Animais , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Meios de Cultura/química , Células Endoteliais/metabolismo , Técnicas In Vitro , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Camundongos
12.
Methods Mol Biol ; 2319: 137-141, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34331251

RESUMO

Lymphatic muscle cells (LMCs), with unique characteristics resembling a combination of both cardiac and smooth muscle cells, play an essential role in the spontaneous contraction of the lymphatic vessels to pump fluid forward. However, our understanding of the more detailed molecular phenotypes of LMCs is limited. Here, we described a method to isolate the LMCs from rat mesentery and then culture the cells in vitro, which will serve a lot more molecular biology study of LMCs and significantly improve our knowledge about the unique characteristics of LMCs.


Assuntos
Técnicas de Cultura de Células/métodos , Separação Celular/métodos , Dissecação/métodos , Mesentério/citologia , Células Musculares/citologia , Animais , Imunofluorescência , Células Musculares/metabolismo , Ratos
13.
Methods Mol Biol ; 2319: 143-152, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34331252

RESUMO

Heart disease is one of the leading causes of death in the United States. Isolation and culture adult cardiomyocytes are important for studying cardiomyocyte contractility, heart hypertrophy, and cardiac failure. In contrast to neonatal cardiomyocyte isolation, adult mice cardiomyocytes isolation is challenging due to firm connections among cardiomyocytes through intercalated discs. The availability of newly generated genetically modified mouse lines requires to establish protocols to isolation and culture adult mouse cardiomyocyte for in vitro studies. In this manuscript, we described a straightforward method of isolating adult mouse cardiomyocytes using Langendorff perfusion apparatus. Briefly, the hearts were harvested from adult mice and the heart was mounted to Lagendorff apparatus. After perfusion with calcium depletion and collagenase digestion, the left ventricles were minced and filtered. Lastly, the separated cardiomyocytes were treated with CaCl2. The isolated cardiac myocytes can be utilized in a broad range of experiments including screening for drugs.


Assuntos
Técnicas de Cultura de Células/métodos , Separação Celular/instrumentação , Separação Celular/métodos , Preparação de Coração Isolado/instrumentação , Preparação de Coração Isolado/métodos , Miócitos Cardíacos/citologia , Animais , Cálcio/farmacologia , Cloreto de Cálcio/farmacologia , Células Cultivadas , Colagenases/química , Colagenases/farmacologia , Camundongos , Miócitos Cardíacos/metabolismo , Perfusão/métodos
14.
Methods Mol Biol ; 2289: 221-233, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270073

RESUMO

The obstacles to breeding programs in Jatropha are the long reproductive cycle with a juvenile phase that lasts several months, the highly heterozygous nature of the genome, the large canopy size, and self-incompatibility that is a long-term process which requires multiple cycles of self-pollination to achieve complete homozygosity. In vitro plant tissue culture-based tools such as haploids and doubled haploid techniques can increase the selection efficiency, resulting into selection of superior plants with complete homozygosity in one generation. It bypasses the complications of greenhouse field evaluation or off-season generation advancement, which takes about 8-10 generations in traditional breeding with the time line of 10-12 years. The haploids have in fact a single set of chromosomes, which undergoes duplication spontaneously during in vitro culture conditions, and are further converted into doubled haploid plants. This represents a major biotechnological tool to accelerate plant breeding. Here, we have established a reproducible, unique anther culture protocol in Jatropha curcas to develop haploid and doubled haploid plants.


Assuntos
Técnicas de Cultura de Células/métodos , Flores/genética , Jatropha/genética , Melhoramento Vegetal/métodos , Árvores/genética , Cromossomos de Plantas/genética , Haploidia , Homozigoto , Polinização/genética
15.
Methods Mol Biol ; 2289: 237-248, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270074

RESUMO

Borage (Borago officinalis L.) is a crop with different culinary, pharmaceutical, and industrial properties. Besides, it is one of the best known sources of gamma linolenic acid (GLA). However, the variability in the levels of such active compounds, obtained from wild borage, may result in conflicting clinical trial reports, which may likely decrease the optimal efficiency of the product. On the other hand, this important medicinal plant has a multifactorial self-incompatibility system, which makes self-pollination ineffective and results in a limited production of pure (homozygous) lines for breeding programs. To avoid the limitations of self-incompatibility and also producing uniform lines useful as parents for F1 hybrid production, or as starting materials to develop new varieties with high and homogenous levels of medicinal compounds, androgenic doubled haploid (DH) lines produced by anther culture have the potential to speed up the process of producing homozygous lines for breeding program of this medicinal species. In the present chapter, a protocol for production of haploid plants in borage by in vitro anther culture is described.


Assuntos
Borago/genética , Técnicas de Cultura de Células/métodos , Flores/genética , Melhoramento Vegetal/métodos , Haploidia , Polinização/genética , Sementes/genética
16.
Methods Mol Biol ; 2289: 249-261, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270075

RESUMO

African violet (Saintpaulia ionantha) is an herbaceous perennial of the Gesneriaceae family. Because almost all the cultivars are heterozygous, pure lines are useful for both classical and new breeding approaches. A shortcut to obtain purebred lines involves the production of doubled haploid strains produced from anther-derived haploids. In this chapter, a protocol for culturing African violet anthers is described in detail.


Assuntos
Técnicas de Cultura de Células/métodos , Flores/genética , Magnoliopsida/genética , Melhoramento Vegetal/métodos , Regeneração/genética , Meios de Cultura/metabolismo , Haploidia
17.
Methods Mol Biol ; 2289: 271-287, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270077

RESUMO

Homozygous parental lines are indispensable for commercial hybrid seed production in many ornamental and vegetable crops. The in vitro induction of haploids and doubled haploids (DHs) through gametic embryogenesis is an effective approach for single-step development of complete homozygous lines from heterozygous donor plants. Anther culture is one of the most popular and widely employed techniques for development of haploids. Here we describe the detailed protocol for rapid and successful induction of haploids in Tagetes spp. using in vitro androgenesis approach. In this protocol, we have provided the comprehensive details of various steps of anther culture in marigold right from the growing of donor plants, selection of buds, pretreatment, embryogenesis and regeneration to ploidy analysis, and chromosome doubling for development of DHs.


Assuntos
Técnicas de Cultura de Células/métodos , Flores/genética , Tagetes/genética , Cromossomos de Plantas/genética , Produtos Agrícolas/genética , Haploidia , Sementes/genética
18.
Methods Mol Biol ; 2289: 289-299, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34270078

RESUMO

Doubled haploid technology allows for producing completely homozygous plants in one generation, which is a very efficient and fast method compared to the production of near-homozygous lines by selfing through conventional breeding methods. However, grain legumes are known to be recalcitrant for most of the in vitro approaches to doubled haploidy. In the last years, significant advances have been made with several legume species through in vitro methods. Chickpea is one of the most important legume species. Several reports have documented the successful generation of haploid plants through anther culture. These reports also showed that successful production of chickpea haploids was achieved when time- and labor-consuming physical stresses such as centrifugation and electroporation were applied to anthers as a pretreatment. In this chapter, we present an efficient and simple anther culture protocol for production of chickpea haploid plants using high concentrations of 2,4-D and silver nitrate in the culture medium, but without applying any physical stresses to anthers.


Assuntos
Técnicas de Cultura de Células/métodos , Cicer/genética , Flores/genética , Melhoramento Vegetal/métodos , Meios de Cultura/metabolismo , Fabaceae/genética , Haploidia , Estresse Fisiológico/genética
19.
Nat Commun ; 12(1): 4492, 2021 07 23.
Artigo em Inglês | MEDLINE | ID: mdl-34301945

RESUMO

Human pluripotent stem cell (hPSC)-derived organoids and cells have similar characteristics to human organs and tissues. Thus, in vitro human organoids and cells serve as a superior alternative to conventional cell lines and animal models in drug development and regenerative medicine. For a simple and reproducible analysis of the quality of organoids and cells to compensate for the shortcomings of existing experimental validation studies, a quantitative evaluation method should be developed. Here, using the GTEx database, we construct a quantitative calculation system to assess similarity to the human organs. To evaluate our system, we generate hPSC-derived organoids and cells, and detected organ similarity. To facilitate the access of our system by researchers, we develop a web-based user interface presenting similarity to the appropriate organs as percentages. Thus, this program could provide valuable information for the generation of high-quality organoids and cells and a strategy to guide proper lineage-oriented differentiation.


Assuntos
Algoritmos , Diferenciação Celular/genética , Especificidade de Órgãos/genética , Organoides/metabolismo , Células-Tronco Pluripotentes/metabolismo , Transcriptoma/genética , Técnicas de Cultura de Células/métodos , Linhagem Celular , Perfilação da Expressão Gênica/métodos , Humanos , Organoides/citologia , Células-Tronco Pluripotentes/citologia , RNA-Seq/métodos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
20.
Int J Mol Sci ; 22(11)2021 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-34198821

RESUMO

Photo-polymerized hydrogels are ideally suited for stem-cell based tissue regeneration and three dimensional (3D) bioprinting because they can be highly biocompatible, injectable, easy to use, and their mechanical and physical properties can be controlled. However, photo-polymerization involves the use of potentially toxic photo-initiators, exposure to ultraviolet light radiation, formation of free radicals that trigger the cross-linking reaction, and other events whose effects on cells are not yet fully understood. The purpose of this study was to examine the effects of hydrogen sulfide (H2S) in mitigating cellular toxicity of photo-polymerization caused to resident cells during the process of hydrogel formation. H2S, which is the latest discovered member of the gasotransmitter family of gaseous signalling molecules, has a number of established beneficial properties, including cell protection from oxidative damage both directly (by acting as a scavenger molecule) and indirectly (by inducing the expression of anti-oxidant proteins in the cell). Cells were exposed to slow release H2S treatment using pre-conditioning with glutathione-conjugated-garlic extract in order to mitigate toxicity during the photo-polymerization process of hydrogel formation. The protective effects of the H2S treatment were evaluated in both an enzymatic model and a 3D cell culture system using cell viability as a quantitative indicator. The protective effect of H2S treatment of cells is a promising approach to enhance cell survival in tissue engineering applications requiring photo-polymerized hydrogel scaffolds.


Assuntos
Técnicas de Cultura de Células/métodos , Hidrogéis/farmacologia , Sulfeto de Hidrogênio/farmacologia , Engenharia Tecidual , Sobrevivência Celular/efeitos dos fármacos , Humanos , Luz , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos da radiação , Polimerização/efeitos dos fármacos , Polimerização/efeitos da radiação , Impressão Tridimensional , Tecidos Suporte , Cicatrização/efeitos dos fármacos , Cicatrização/efeitos da radiação
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...