It is now widely recognized that mesenchymal stem cells (MSCs) possess the capacity to differentiate into a wide array of cell types. Numerous studies have identified the role of lncRNA in the regulation of MSC differentiation. It is important to elucidate the role and interplay of microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) in the regulation of signalling pathways that govern MSC function. Furthermore, miRNAs and lncRNAs are important clinical for innovative strategies aimed at addressing a wide spectrum of existing and emerging disease. Hence it is important to consider their impact on MSC function and differentiation. Examining the data available in public databases, we have collected the literature containing the latest discoveries pertaining to human stem cells and their potential in both fundamental research and clinical applications. Furthermore, we have compiled completed clinical studies that revolve around the application of MSCs, shedding light on the opportunities presented by harnessing the regulatory potential of miRNAs and lncRNAs. This exploration of the therapeutic possibilities offered by miRNAs and lncRNAs within MSCs unveils exciting prospects for the development of precision therapies and personalized treatment approaches. Ultimately, these advancements promise to augment the efficacy of regenerative strategies and produce positive outcomes for patients. As research in this field continues to evolve, it is imperative to explore and exploit the vast potential of miRNAs and lncRNAs as therapeutic agents. The findings provide a solid basis for ongoing investigations, fuelling the quest to fully unlock the regenerative potential of MSCs.
- Klíčová slova
- lncRNA, mesenchymal stem cells, miRNA,
- MeSH
- buněčná diferenciace genetika MeSH
- lidé MeSH
- mezenchymální kmenové buňky * metabolismus MeSH
- mikro RNA * genetika metabolismus MeSH
- osteoblasty metabolismus MeSH
- RNA dlouhá nekódující * genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- mikro RNA * MeSH
- RNA dlouhá nekódující * MeSH
Healing of dense regular connective tissue, due to a high fiber-to-cell ratio and low metabolic activity and regeneration potential, frequently requires surgical implantation or reconstruction with high risk of reinjury. An alternative to synthetic implants is using bioscaffolds obtained through decellularization, a process where the aim is to extract cells from the tissue while preserving the tissue-specific native molecular structure of the ECM. Proteins, lipids, nucleic acids and other various extracellular molecules are largely involved in differentiation, proliferation, vascularization and collagen fibers deposit, making them the crucial processes in tissue regeneration. Because of the multiple possible forms of cell extraction, there is no standardized protocol in dense regular connective tissue (DRCT). Many modifications of the structure, shape and composition of the bioscaffold have also been described to improve the therapeutic result following the implantation of decellularized connective tissue. The available data provide a valuable source of crucial information. However, the wide spectrum of decellularization makes it important to understand the key aspects of bioscaffolds relative to their potential use in tissue regeneration.
- Klíčová slova
- bioscaffold, extracellular matrix, ligament, tendon,
- MeSH
- buněčná diferenciace MeSH
- implantace embrya MeSH
- lidé MeSH
- nukleové kyseliny * MeSH
- patologická angiogeneze MeSH
- regenerativní lékařství * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- nukleové kyseliny * MeSH
Exosomal regulation is intimately involved in key cellular processes, such as migration, proliferation, and adhesion. By participating in the regulation of basic mechanisms, extracellular vesicles are important in intercellular signaling and the functioning of the mammalian reproductive system. The complexity of intercellular interactions in the ovarian follicle is also based on multilevel intercellular signaling, including the mechanisms involving cadherins, integrins, and the extracellular matrix. The processes in the ovary leading to the formation of a fertilization-ready oocyte are extremely complex at the molecular level and depend on the oocyte's ongoing relationship with granulosa cells. An analysis of gene expression from material obtained from a primary in vitro culture of porcine granulosa cells was employed using microarray technology. Genes with the highest expression (LIPG, HSD3B1, CLIP4, LOX, ANKRD1, FMOD, SHAS2, TAGLN, ITGA8, MXRA5, and NEXN) and the lowest expression levels (DAPL1, HSD17B1, SNX31, FST, NEBL, CXCL10, RGS2, MAL2, IHH, and TRIB2) were selected for further analysis. The gene expression results obtained from the microarrays were validated using quantitative RT-qPCR. Exosomes may play important roles regarding intercellular signaling between granulosa cells. Therefore, exosomes may have significant applications in regenerative medicine, targeted therapy, and assisted reproduction technologies.
- Klíčová slova
- cell adhesion, cell migration and proliferation, cellular signaling, extracellular matrix, extracellular vesicles, porcine granulosa cells, transcriptomics,
- MeSH
- folikulární buňky * metabolismus MeSH
- oocyty metabolismus MeSH
- ovariální folikul * metabolismus MeSH
- ovarium metabolismus MeSH
- prasata MeSH
- proliferace buněk genetika MeSH
- savci MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The ovarian follicle is the basic functional unit of the ovary, comprising theca cells and granulosa cells (GCs). Two different types of GCs, mural GCs and cumulus cells (CCs), serve different functions during folliculogenesis. Mural GCs produce oestrogen during the follicular phase and progesterone after ovulation, while CCs surround the oocyte tightly and form the cumulus oophurus and corona radiata inner cell layer. CCs are also engaged in bi-directional metabolite exchange with the oocyte, as they form gap-junctions, which are crucial for both the oocyte's proper maturation and GC proliferation. However, the function of both GCs and CCs is dependent on proper follicular angiogenesis. Aside from participating in complex molecular interplay with the oocyte, the ovarian follicular cells exhibit stem-like properties, characteristic of mesenchymal stem cells (MSCs). Both GCs and CCs remain under the influence of various miRNAs, and some of them may contribute to polycystic ovary syndrome (PCOS) or premature ovarian insufficiency (POI) occurrence. Considering increasing female fertility problems worldwide, it is of interest to develop new strategies enhancing assisted reproductive techniques. Therefore, it is important to carefully consider GCs as ovarian stem cells in terms of the cellular features and molecular pathways involved in their development and interactions as well as outline their possible application in translational medicine.
- Klíčová slova
- cumulus cells, follicular angiogenesis, granulosa cells, miRNA, stem cells, translational medicine,
- MeSH
- fyziologická neovaskularizace * MeSH
- kmenové buňky metabolismus MeSH
- kumulární buňky metabolismus MeSH
- lidé MeSH
- primární ovariální insuficience metabolismus MeSH
- syndrom polycystických ovarií metabolismus MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Adult myogenesis is dependent on a population of precursor cells, located between the sarcolemma and the basal lamina of the muscle fiber. These satellite cells, usually present in a quiescent state, become activated in response to mechanical muscle strain, differentiating and fusing to add new nuclei to enlarging muscles. As their myogenic lineage commitment is induced on demand, muscle satellite cells exhibit a certain amount of plasticity, possibly being able to be directed to differentiate into non-myogenic fates. In this study, myosatellite cells were isolated from chicken muscle samples, characterized in vitro and introduced into developing blastoderms. They were further investigated using fluorescence microscopy, immunohistochemistry and PCR, to determine their location in embryos after three and eighteen days. The results of the in vitro analysis confirmed that the cells obtained from the Pectoralis thoracicus are highly myogenic, based on the expression of Pax7, Myogenin, MyoD, Desmin and the myotube assay. Furthermore, the investigation of satellite cells within the embryo showed their migration to the regions of Pectoralis thoracicus, heart, liver, gizzard, proventriculus, intestine and brain. Overall, the results of the study proved the high myogenicity of chicken Pectoralis thoracicus cell isolates, as well as provided new information about their migration pathways following introduction into the blastocyst. The presence of the introduced LacZ or eGFP transgenes across the embryo, even 20 days after myosatellite cell injection, further supports the notion that satellite cells exhibit significant plasticity, potentially transdifferentiating into non-muscle lineages.
- Klíčová slova
- chicken, embryo, plasticity, satellite cells, stem cells,
- Publikační typ
- časopisecké články MeSH
Ammonia is very toxic to the body and has detrimental effects on many different organ systems. Using cultured myoblast cells, we examined ammonia's effect on myostatin expression, a negative regulator of skeletal muscle growth, and myotube diameters. The objective of this study was to examine how murine, avian, and fish cells respond to increasing levels of ammonia up to 50 mM. The murine myoblast cell line (C2C12), primary chick, and primary tilapia myoblast cells were cultured and then exposed to 10, 25, and 50 mM ammonium acetate, sodium acetate, and an untreated control for 24 h. High levels of ammonia were detrimental to the C2C12 cells, causing increased Myostatin (MSTN) expression and decreased myotube diameters between 10 and 25 mM (p < 0.002). Ammonia at 10 mM continued the positive myogenic response in the chick, with lower MSTN expression than the C2C12 cells and larger myotube diameters, but the myotube diameter at 50 mM ammonium acetate was significantly smaller than those at 10 and 25 mM (p < 0.001). However, chick myotubes at 50 mM were still significantly larger than the sodium acetate-treated and untreated control (p < 0.001). The tilapia cells showed no significant difference in MSTN expression or myotube diameter in response to increasing the concentrations of ammonia. Overall, these results confirm that increasing concentrations of ammonia are detrimental to mammalian skeletal muscle, while chick cells responded positively at lower levels but began to exhibit a negative response at higher levels, as the tilapia experienced no detrimental effects. The differences in ammonia metabolism strategies between fish, avian, and mammalian species could potentially contribute to the differences between species in response to high levels of ammonia. Understanding how ammonia affects skeletal muscle is important for the treatment of muscle wasting observed in liver failure patients.
- Klíčová slova
- ammonia, avian, fish, mammals, muscle, myostatin,
- MeSH
- amoniak farmakologie MeSH
- buněčná diferenciace * MeSH
- kosterní svaly účinky léků metabolismus patologie MeSH
- lidé MeSH
- myostatin genetika metabolismus MeSH
- myši MeSH
- ptáci MeSH
- regulace genové exprese účinky léků MeSH
- ryby MeSH
- vývoj svalů * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- amoniak MeSH
- myostatin MeSH
Polyspermia is an adverse phenomenon during mammalian fertilization when more than one sperm fuses with a single oocyte. The egg cell is prepared to prevent polyspermia by, among other ways, producing cortical granules (CGs), which are specialized intracellular structures containing enzymes that aim to harden the zona pellucida and block the fusion of subsequent sperm. This work focused on exploring the expression profile of genes that may be associated with cortical reactions, and evaluated the distribution of CGs in immature oocytes and the peripheral density of CGs in mature oocytes. Oocytes were isolated and then processed for in vitro maturation (IVM). Transcriptomic analysis of genes belonging to five ontological groups has been conducted. Six genes showed increased expression after IVM (ARHGEF2, MAP1B, CXCL12, FN1, DAB2, and SOX9), while the majority of genes decreased expression after IVM. Using CG distribution analysis in immature oocytes, movement towards the cortical zone of the oocyte during meiotic competence acquisition was observed. CGs peripheral density decreased with the rise in meiotic competence during the IVM process. The current results reveal important new insights into the in vitro maturation of oocytes. Our results may serve as a basis for further studies to investigate the cortical reaction of oocytes.
- Klíčová slova
- cortical granule, microarray, molecular markers, oocyte maturation, pig,
- MeSH
- buněčná diferenciace * MeSH
- cytoplazmatická granula metabolismus MeSH
- IVM techniky metody MeSH
- kultivované buňky MeSH
- oocyty cytologie metabolismus MeSH
- prasata MeSH
- transkriptom * MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The primary function of ovarian granulosa cells (GCs) is the support of oocytes during maturation and development. Molecular analyses of granulosa cell-associated processes, leading to improvement of understanding of the cell cycle events during the formation of ovarian follicles (folliculogenesis), may be key to improve the in vitro fertilization procedures. Primary in vitro culture of porcine GCs was employed to examine the changes in the transcriptomic profile of genes belonging to "cell cycle", "cell division", "cell cycle process", "cell cycle phase transition", "cell cycle G1/S phase transition", "cell cycle G2/M phase transition" and "cell cycle checkpoint" ontology groups. During the analysis, microarrays were employed to study the transcriptome of GCs, analyzing the total RNA of cells from specific periods of in vitro cultures. This research was based on material obtained from 40 landrace gilts of similar weight, age and the same living conditions. RNA was isolated at specific timeframes: before the culture was established (0 h) and after 48 h, 96 h and 144 h in vitro. Out of 133 differentially expressed genes, we chose the 10 most up-regulated (SFRP2, PDPN, PDE3A, FGFR2, PLK2, THBS1, ETS1, LIF, ANXA1, TGFB1) and the 10 most downregulated (IGF1, NCAPD2, CABLES1, H1FOO, NEK2, PPAT, TXNIP, NUP210, RGS2 and CCNE2). Some of these genes known to play key roles in the regulation of correct cell cycle passage (up-regulated SFRP2, PDE3A, PLK2, LIF and down-regulated CCNE2, TXNIP, NEK2). The data obtained provide a potential reference for studies on the process of mammalian folliculogenesis, as well as suggests possible new genetic markers for cell cycle progress in in vitro cultured porcine granulosa cells.
- Klíčová slova
- Granulosa cells, Microarray, Ovarian follicle, Pig, Primary culture,
- MeSH
- buněčný cyklus genetika MeSH
- folikulární buňky cytologie MeSH
- kultivované buňky MeSH
- ovariální folikul cytologie MeSH
- prasata MeSH
- stanovení celkové genové exprese MeSH
- transkriptom * MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH