Efficient and high yield isolation of myoblasts from skeletal muscle

Shahini, Aref; Vydiam, Kalyan; Choudhury, Debanik; Rajabian, Nika; Nguyen, T M; Lei, Pedro; Andreadis, Stelios T.

doi:10.1016/j.scr.2018.05.017

Cited by 82 publications

(74 citation statements)

References 36 publications

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“…Moreover, further myosatellite markers were examined in cells obtained from the 1-day-old chicks: MyoD and myogenin, as well as known myogenic regulatory factors and desmin, which is a generally accepted marker of satellite cells after three days in culture [19,26,27]. Desmin was detected in 100% of the isolated cells, while the levels of MyoD, myogenin and Pax7 reflected previously reported in vivo levels of cells expressing the three markers [28].…”

Section: Discussionmentioning

confidence: 60%

Avian Satellite Cell Plasticity

Jankowski

Mozdziak

Petitte

et al. 2020

Animals

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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.

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Section: Discussionmentioning

confidence: 60%

Avian Satellite Cell Plasticity

Jankowski

Mozdziak

Petitte

et al. 2020

Animals

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“…It has also been shown that 95% of the preplated rat muscle cells from the hind limb and back were positive for desmin and MyoD (Sheehan, Tatsumi, Temm‐Grove, & Allen, 2000). The purity of mouse myoblasts was increased up to 98% through preplating based on the expression of myogenic markers such as integrin α7, MyoD, and desmin (Shahini et al., 2018). Moreover, Qu‐Petersen et al.…”

Section: Muscle Stem Cell Isolation From Tissuesmentioning

confidence: 99%

“…The stemness of the cells significantly increased in a dose‐dependent manner under these conditions. Furthermore, high serum‐containing media with 20% FBS and 10% HS exclusively promote mouse muscle stem cell proliferation compared with that of nonmyogenic cells, especially fibroblasts (Shahini et al., 2018; Wang et al., 2014). Currently, due to the donor animal‐based wide variations in their composition, serum replacements, such as knockout serum replacement (KSR), and N2/B27 supplements are used to obtain consistent experimental results.…”

Section: In Vitro Culture Of Muscle Stem Cellsmentioning

confidence: 99%

Muscle stem cell isolation and in vitro culture for meat production: A methodological review

Choi

Yoon

Kim

et al. 2020

Comp Rev Food Sci Food Safe

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Cultured muscle tissue‐based protein products, also known as cultured meat, are produced through in vitro myogenesis involving muscle stem cell culture and differentiation, and mature muscle cell processing for flavor and texture. This review focuses on the in vitro myogenesis for cultured meat production. The muscle stem cell–based in vitro muscle tissue production consists of a sequential process: (1) muscle sampling for stem cell collection, (2) muscle tissue dissociation and muscle stem cell isolation, (3) primary cell culture, (4) upscaled cell culture, (5) muscle differentiation and maturation, and (6) muscle tissue harvest. Although muscle stem cell research is a well‐established field, the majority of these steps remain to be underoptimized to enable the in vitro creation of edible muscle‐derived meat products. The profound understanding of the process would help not only cultured meat production but also business sectors that have been seeking new biomaterials for the food industry. In this review, we discuss comprehensively and in detail each step of cutting‐edge methods for cultured meat production. This would be meaningful for both academia and industry to prepare for the new era of cellular agriculture.

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“…In order to generate implantable neuromuscular constructs with anatomical relevance, we propose to stretch-grow engineered motor neuron aggregates on a bed of pre-differentiated myofibers grown on a suitable substrate 166 . Motor neurons can be harvested from rodent spinal cords 182 or obtained from differentiated human PSCs 183 using published protocols, while skeletal muscle cells can be sourced from cell lines, rodents 184 , or human PSCs 185 . This concept would entail culturing the skeletal muscle cells on a scaffold for 4-7 days in differentiation media to allow for the formation of myofibers prior to introduction and mechanical stretch of motor neurons in mechanobioreactors.…”

Section: Pre-innervated Skeletal Musclementioning

confidence: 99%

Innervation: the missing link for biofabricated tissues and organs

et al. 2020

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Innervation plays a pivotal role as a driver of tissue and organ development as well as a means for their functional control and modulation. Therefore, innervation should be carefully considered throughout the process of biofabrication of engineered tissues and organs. Unfortunately, innervation has generally been overlooked in most non-neural tissue engineering applications, in part due to the intrinsic complexity of building organs containing heterogeneous native cell types and structures. To achieve proper innervation of engineered tissues and organs, specific host axon populations typically need to be precisely driven to appropriate location(s) within the construct, often over long distances. As such, neural tissue engineering and/or axon guidance strategies should be a necessary adjunct to most organogenesis endeavors across multiple tissue and organ systems. To address this challenge, our team is actively building axon-based "living scaffolds" that may physically wire in during organ development in bioreactors and/or serve as a substrate to effectively drive targeted long-distance growth and integration of host axons after implantation. This article reviews the neuroanatomy and the role of innervation in the functional regulation of cardiac, skeletal, and smooth muscle tissue and highlights potential strategies to promote innervation of biofabricated engineered muscles, as well as the use of "living scaffolds" in this endeavor for both in vitro and in vivo applications. We assert that innervation should be included as a necessary component for tissue and organ biofabrication, and that strategies to orchestrate host axonal integration are advantageous to ensure proper function, tolerance, assimilation, and bio-regulation with the recipient post-implant.

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Efficient and high yield isolation of myoblasts from skeletal muscle

Cited by 82 publications

References 36 publications

Avian Satellite Cell Plasticity

Avian Satellite Cell Plasticity

Muscle stem cell isolation and in vitro culture for meat production: A methodological review

Innervation: the missing link for biofabricated tissues and organs

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