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Food Research International (Ottawa,... Oct 2023Cultivated meat production requires an efficient, robust and highly optimized serum-free cell culture media for the needed upscaling of muscle cell expansion. Existing...
Cultivated meat production requires an efficient, robust and highly optimized serum-free cell culture media for the needed upscaling of muscle cell expansion. Existing formulations of serum-free media are complex, expensive and have not been optimized for muscle cells. Thus, we undertook this work to develop a simple and robust serum-free media for the proliferation of bovine satellite cells (SCs) through Design of Experiment (DOE) and Response Surface Methodology (RSM) using precise and high-throughput image-based cytometry. Proliferative attributes were investigated with transcriptomics and long-term performance was validated using multiple live assays. Here we formulated a media based on three highly optimized components; FGF2 (2 ng/mL), fetuin (600 µg/mL) and BSA (75 µg/mL) which together with an insulin-transferrin-selenium (1x) supplement, sustained the proliferation of bovine SCs, porcine SCs and murine C2C12 muscle cells. Remarkably, cells cultured in our media named Tri-basal 2.0+ performed better than cell cultured in 10% FBS, with respect to proliferation. Hence, the optimized Tri-basal 2.0+ enhanced serum-free cell attachment and long-term proliferation, providing an alternative solution to the use of FBS in the production of cultivated meat.
Topics: Animals; Cattle; Mice; Swine; Culture Media, Serum-Free; Muscle Cells; Muscles; Biological Assay; Cell Proliferation
PubMed: 37689947
DOI: 10.1016/j.foodres.2023.113194 -
Current Topics in Developmental Biology 2024Myocyte fusion is a pivotal process in the development and regeneration of skeletal muscle. Failure during fusion can lead to a range of developmental as well as... (Review)
Review
Myocyte fusion is a pivotal process in the development and regeneration of skeletal muscle. Failure during fusion can lead to a range of developmental as well as pathological consequences. This review aims to comprehensively explore the intricate processes underlying myocyte fusion, from the molecular to tissue scale. We shed light on key players, such as the muscle-specific fusogens - Myomaker and Myomixer, in addition to some lesser studied molecules contributing to myocyte fusion. Conserved across vertebrates, Myomaker and Myomixer play a crucial role in driving the merger of plasma membranes of fusing myocytes, ensuring the formation of functional muscle syncytia. Our multiscale approach also delves into broader cell and tissue dynamics that orchestrate the timing and positioning of fusion events. In addition, we explore the relevance of muscle fusogens to human health and disease. Mutations in fusogen genes have been linked to congenital myopathies, providing unique insights into the molecular basis of muscle diseases. We conclude with a discussion on potential therapeutic avenues that may emerge from manipulating the myocyte fusion process to remediate skeletal muscle disorders.
Topics: Humans; Cell Fusion; Animals; Muscle, Skeletal; Muscle Cells; Muscle Proteins
PubMed: 38670716
DOI: 10.1016/bs.ctdb.2024.01.016 -
Reproduction (Cambridge, England) Mar 2020The oviduct (known as the fallopian tube in humans) is the site for fertilization and pre-implantation embryo development. Female steroid hormones, estrogen and... (Review)
Review
The oviduct (known as the fallopian tube in humans) is the site for fertilization and pre-implantation embryo development. Female steroid hormones, estrogen and progesterone, are known to modulate the morphology and function of cells in the oviduct. In this review, we focus on the actions of estrogen and progesterone on secretory, ciliated, and muscle cell functions and morphologies during fertilization, pre-implantation embryo development, and embryo transport in humans, laboratory rodents and farm animals. We review some aspects of oviductal anatomy and histology and discuss current assisted reproductive technologies (ARTs) that bypass the oviduct and their effects on embryo quality. Lastly, we review the causes of alterations in secretory, ciliated, and muscle cell functions that could result in embryo transport defects.
Topics: Animals; Animals, Domestic; Embryonic Development; Epithelial Cells; Estrogens; Fallopian Tubes; Female; Genital Diseases, Female; Humans; Menstrual Cycle; Mice; Microscopy, Electron, Scanning; Muscle Cells; Pregnancy; Pregnancy, Ectopic; Progesterone; Rats; Reproductive Techniques, Assisted
PubMed: 32040278
DOI: 10.1530/REP-19-0189 -
Cell Death & Disease Dec 2023Lipotoxicity, the accumulation of lipids in non-adipose tissues, alters the metabolic transcriptome and mitochondrial metabolism in skeletal muscle. The mechanisms...
Lipotoxicity, the accumulation of lipids in non-adipose tissues, alters the metabolic transcriptome and mitochondrial metabolism in skeletal muscle. The mechanisms involved remain poorly understood. Here we show that lipotoxicity increased histone deacetylase 4 (HDAC4) and histone deacetylase 5 (HDAC5), which reduced the expression of metabolic genes and oxidative metabolism in skeletal muscle, resulting in increased non-oxidative glucose metabolism. This metabolic reprogramming was also associated with impaired apoptosis and ferroptosis responses, and preserved muscle cell viability in response to lipotoxicity. Mechanistically, increased HDAC4 and 5 decreased acetylation of p53 at K120, a modification required for transcriptional activation of apoptosis. Redox drivers of ferroptosis derived from oxidative metabolism were also reduced. The relevance of this pathway was demonstrated by overexpression of loss-of-function HDAC4 and HDAC5 mutants in skeletal muscle of obese db/db mice, which enhanced oxidative metabolic capacity, increased apoptosis and ferroptosis and reduced muscle mass. This study identifies HDAC4 and HDAC5 as repressors of skeletal muscle oxidative metabolism, which is linked to inhibition of cell death pathways and preservation of muscle integrity in response to lipotoxicity.
Topics: Mice; Animals; Histone Deacetylases; Muscle Cells; Muscle, Skeletal; Protein Processing, Post-Translational; Cell Death
PubMed: 38040704
DOI: 10.1038/s41419-023-06319-5 -
Handbook of Experimental Pharmacology 2022In humans, smooth muscle cells (SMCs) are the main cell type in the artery medial layer, in pre-atherosclerotic diffuse thickening of the intima, and in all stages of... (Review)
Review
In humans, smooth muscle cells (SMCs) are the main cell type in the artery medial layer, in pre-atherosclerotic diffuse thickening of the intima, and in all stages of atherosclerotic lesion development. SMCs secrete the proteoglycans responsible for the initial binding and retention of atherogenic lipoproteins in the artery intima, with this retention driving foam cell formation and subsequent stages of atherosclerosis. In this chapter we review current knowledge of the extracellular matrix generated by SMCs in medial and intimal arterial layers, their relationship to atherosclerotic lesion development and stabilization, how these findings correlate with mouse models of atherosclerosis, and potential therapies aimed at targeting the SMC matrix-lipoprotein interaction for atherosclerosis prevention.
Topics: Animals; Atherosclerosis; Lipoproteins; Mice; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Proteoglycans
PubMed: 33340050
DOI: 10.1007/164_2020_364 -
Neuromuscular Disorders : NMD Jun 2020Emery-Dreifuss muscular dystrophy (EDMD) is a rare genetic disorder characterised by the early development of muscle contractures, progressive muscle weakness, and heart... (Review)
Review
Emery-Dreifuss muscular dystrophy (EDMD) is a rare genetic disorder characterised by the early development of muscle contractures, progressive muscle weakness, and heart abnormalities. The latter may result in serious complications, or in severe cases, sudden death. Currently, there are very few effective treatment options available for EDMD and so there is a high clinical need for new therapies. Various genetic mutations have been identified in the development and causation of EDMD, each encoding proteins that are components of the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex, which spans the nuclear envelope and serves to connect the nuclear lamina to the cytoskeleton. Within this review, we examine how mutations in the genes encoding these proteins, including lamins A/C, emerin, nesprins 1/2, FHL1, and SUN1/2 lead to muscle cell differentiation and development pathway defects. Further work to identify conserved molecular pathways downstream of these defective proteins may reveal potential targets for therapy design.
Topics: Animals; Cell Differentiation; Gene Expression Regulation, Developmental; Humans; Membrane Proteins; Muscle Cells; Muscle Proteins; Muscular Dystrophy, Emery-Dreifuss; Nuclear Proteins; Signal Transduction
PubMed: 32522500
DOI: 10.1016/j.nmd.2020.04.002 -
Cellular Signalling Jun 2020Abdominal aortic aneurysms (AAAs) are highly lethal cardiovascular diseases without effective medications. However, the molecular and signaling mechanisms remain... (Review)
Review
Abdominal aortic aneurysms (AAAs) are highly lethal cardiovascular diseases without effective medications. However, the molecular and signaling mechanisms remain unclear. A series of pathological cellular processes have been shown to contribute to AAA formation, including vascular extracellular matrix remodeling, inflammatory and immune responses, oxidative stress, and dysfunction of vascular smooth muscle cells. Each cellular process involves complex cellular signaling, such as NF-κB, MAPK, TGFβ, Notch and inflammasome signaling. In this review, we discuss how cellular signaling networks function in various cellular processes during the pathogenesis and progression of AAA. Understanding the interaction of cellular signaling networks with AAA pathogenesis as well as the crosstalk of different signaling pathways is essential for the development of novel therapeutic approaches to and personalized treatments of AAA diseases.
Topics: Animals; Aortic Aneurysm, Abdominal; Humans; Myocytes, Smooth Muscle; Oxidative Stress; Signal Transduction; Vascular Remodeling
PubMed: 32088371
DOI: 10.1016/j.cellsig.2020.109575 -
International Journal of Oral Science Jun 2023Periodontitis imparting the increased risk of atherosclerotic cardiovascular diseases is partially due to the immune subversion of the oral pathogen, particularly the...
Periodontitis imparting the increased risk of atherosclerotic cardiovascular diseases is partially due to the immune subversion of the oral pathogen, particularly the Porphyromonas gingivalis (P. gingivalis), by inducing apoptosis. However, it remains obscure whether accumulated apoptotic cells in P. gingivalis-accelerated plaque formation are associated with impaired macrophage clearance. Here, we show that smooth muscle cells (SMCs) have a greater susceptibility to P. gingivalis-induced apoptosis than endothelial cells through TLR2 pathway activation. Meanwhile, large amounts of miR-143/145 in P.gingivalis-infected SMCs are extracellularly released and captured by macrophages. Then, these miR-143/145 are translocated into the nucleus to promote Siglec-G transcription, which represses macrophage efferocytosis. By constructing three genetic mouse models, we further confirm the in vivo roles of TLR2 and miR-143/145 in P. gingivalis-accelerated atherosclerosis. Therapeutically, we develop P.gingivalis-pretreated macrophage membranes to coat metronidazole and anti-Siglec-G antibodies for treating atherosclerosis and periodontitis simultaneously. Our findings extend the knowledge of the mechanism and therapeutic strategy in oral pathogen-associated systemic diseases.
Topics: Animals; Mice; Endothelial Cells; Toll-Like Receptor 2; Macrophages; Apoptosis; Atherosclerosis; Myocytes, Smooth Muscle; MicroRNAs
PubMed: 37380627
DOI: 10.1038/s41368-023-00232-5 -
Journal of Healthcare Engineering 2022Atherosclerosis is a chronic inflammatory disease of the arterial wall and the main cause of cardiovascular disease and cerebrovascular disease. In recent years, the...
Atherosclerosis is a chronic inflammatory disease of the arterial wall and the main cause of cardiovascular disease and cerebrovascular disease. In recent years, the mortality rate of atherosclerotic diseases has become higher and higher. This article aims to study the dysregulation of atherosclerotic vascular endothelial secretion and smooth muscle cell proliferation, and put forward and practice the pathological research of atherosclerotic disease. This article describes in detail atherosclerosis, endothelial dysfunction, and smooth muscle cell proliferation, and studies the causes of atherosclerosis. Research results indicate that atherosclerotic vascular endothelial dysfunction also has a great influence on the proliferation of smooth muscle cells. Many genes and environmental factors can regulate the functions of endothelial cells, vascular smooth muscle cells, and mononuclear macrophages and affect the formation of atherosclerosis. At the same time, diabetes, hypertension, hyperlipidemia, obesity, etc. are the main causes of atherosclerosis. The number of patients with cardiovascular and cerebrovascular diseases dying from atherosclerosis in the country is increasing, and the proportion is close to 30%.
Topics: Atherosclerosis; Cell Proliferation; Endothelial Cells; Humans; Myocytes, Smooth Muscle
PubMed: 35310191
DOI: 10.1155/2022/9271879 -
Scientific Reports Mar 2023Cell-cultivated fish offers the potential for a more ethical, sustainable, and safe seafood system. However, fish cell culture is relatively understudied in comparison...
Cell-cultivated fish offers the potential for a more ethical, sustainable, and safe seafood system. However, fish cell culture is relatively understudied in comparison to mammalian cells. Here, we established and characterized a continuous Atlantic mackerel (Scomber scombrus) skeletal muscle cell line ("Mack" cells). The cells were isolated from muscle biopsies of fresh-caught fish, with separate isolations performed from two distinct fish. Mack1 cells (cells from the first isolation) were cultured for over a year and subcultured over 130 times. The cells proliferated at initial doubling times of 63.9 h (± 19.1 SD). After a spontaneous immortalization crisis from passages 37-43, the cells proliferated at doubling times of 24.3 h (± 4.91 SD). A muscle phenotype was confirmed through characterization of muscle stemness and differentiation via paired-box protein 7 and myosin heavy chain immunostaining, respectively. An adipocyte-like phenotype was also demonstrated for the cells through lipid accumulation, confirmed via Oil Red O staining and quantification of neutral lipids. New qPCR primers (HPRT, PAX3B, MYOD1, MYOG, TNNT3A, and PPARG) were tailored to the mackerel genome and used to characterize mackerel cell genotypes. This work provides the first spontaneously immortalized fish muscle cell line for research, ideally serving as a reference for subsequent investigation.
Topics: Animals; Muscles; Fishes; Perciformes; Muscle Cells; Cell Line; Phenotype; Mammals
PubMed: 36991012
DOI: 10.1038/s41598-023-31822-2