Characterization and isolation of highly purified porcine satellite cells

Ding, Shijie; Wang, Fei; Liu, Yan; Li, Sheng; Zhou, Guanghong; Hu, Ping

doi:10.1038/cddiscovery.2017.3

Cited by 86 publications

(106 citation statements)

References 54 publications

(53 reference statements)

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“…At 40–60 min after seeding on the collagen-coated culture plate, the stem cell population can be obtained by harvesting the supernatant, since most of the fibroblasts and epithelial cells remain attached to the culture plate ( Rando and Blau, 1994 ; Richler and Yaffe, 1970 ). However, density gradient centrifugation and the preplating technique reportedly show wide variations and low fidelities ( Ding et al, 2017 ). Advances in molecular biology allow us to analyze and separate the cells based on their molecular features.…”

Section: Introductionmentioning

confidence: 99%

Purification of Pig Muscle Stem Cells Using Magnetic-Activated Cell Sorting (MACS) Based on the Expression of Cluster of Differentiation 29 (CD29)

Choi¹,

Kim²,

Yoon³

et al. 2020

Food Sci Anim Resour

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The muscle stem cells of domestic animals are of interest to researchers in the food and biotechnology industries for the production of cultured meat. For producing cultured meat, it is crucial for muscle stem cells to be efficiently isolated and stably maintained in vitro on a large scale. In the present study, we aimed to optimize the method for the enrichment of pig muscle stem cells using a magnetic-activated cell sorting (MACS) system. Pig muscle stem cells were collected from the biceps femoris muscles of 14 d-old pigs of three breeds [Landrace×Yorkshire×Duroc (LYD), Berkshire, and Korean native pigs] and cultured in skeletal muscle growth medium-2 (SkGM-2) supplemented with epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580). Approximately 30% of total cultured cells were nonmyogenic cells in the absence of purification in our system, as determined by immunostaining for cluster of differentiation 56 (CD56) and CD29, which are known markers of muscle stem cells. Interestingly, following MACS isolation using the CD29 antibody, the proportion of CD56 + /CD29 + muscle stem cells was significantly increased (91.5±2.40%), and the proportion of CD56 single-positive nonmyogenic cells was dramatically decreased. Furthermore, we verified that this method worked well for purifying muscle stem cells in the three pig breeds. Accordingly, we found that CD29 is a valuable candidate among the various marker genes for the isolation of pig muscle stem cells and developed a simple sorting method based on a single antibody to this protein.

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Section: Introductionmentioning

confidence: 99%

Purification of Pig Muscle Stem Cells Using Magnetic-Activated Cell Sorting (MACS) Based on the Expression of Cluster of Differentiation 29 (CD29)

Choi¹,

Kim²,

Yoon³

et al. 2020

Food Sci Anim Resour

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“…CD56 has been mainly described as a marker for identification of quiescent human MuSCs (Alexander et al, 2016;Castiglioni et al, 2014;Uezumi et al, 2016). CD29, has been previously used for the isolation of MuSCs from mouse, pig, and cow (Ding et al, 2018;Ding et al, 2017;Maesner et al, 2016). It is a member of the integrin family and interacts with both collagen and laminin depending on its heterodimer binding partner (Hynes, 2002).…”

Section: Determination Of Reliable Positive Markers For Rat Musc Isolmentioning

confidence: 99%

“…Isolation of MuSCs has been described in mouse, human, pig, and cow (Alexander et al, 2016;Ding et al, 2018;Ding et al, 2017;Liu et al, 2015;Maesner et al, 2016;Uezumi et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Isolation of muscle stem cells from rat skeletal muscles

Sesillo

Wong

Cortez

et al. 2019

Preprint

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Background: Muscle stem cells (MuSCs) are involved in homeostatic maintenance of skeletal muscles and play a central role in muscle regeneration in response to injury. Thus, understanding MuSC autonomous properties is of fundamental importance for studies of muscle degenerative diseases and muscle plasticity. Rat, as an animal model, has been widely used in the skeletal muscle field, however an efficient approach for MuSC isolation through fluorescence-activated cell sorting from rat muscles has never been described. This work aims to develop and validate an effective protocol for MuSC isolation from rat skeletal muscles.Methods: Tibialis anterior, gastrocnemius, diaphragm, and the individual components of the pelvic floor muscle complex (coccygeus, iliocaudalis, and pubocaudalis) were harvested from female rats and digested for isolation of MuSCs. Three protocols, employing different cell surface markers (CD106, CD56, and CD29), were compared for their ability to isolate a pure MuSC population.Results: Cells obtained using the protocol that relies only on VCAM-1 (CD106) as a positive marker showed high expression of Pax7 upon isolation, ability to progress through myogenic lineage while in culture, and complete differentiation in serum deprived conditions. The protocol was further validated in other skeletal muscles proving to be reproducible. Conclusions:CD106 is an efficient marker for reliable isolation of MuSCs from a variety of rat skeletal muscles.

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“…Myogenesis is the process of skeletal muscle development and formation, which is a highly coordinated developmental process involving the coordinate expression of a number of myogenic regulatory factors, such as paired box 3, paired box 7, myogenic factor 5, myogenin (myog), and myogenic differentiation (MYOD) [1][2][3]. Satellite cells (SCs) reside between basement membranes and muscle sarcolemma in myofibers and play a vital role in the regeneration and differentiation of skeletal muscle [4,5]. SCs are activated by specific signals such as muscle injury, and can self-renew or enter the myogenic pathway and subsequently differentiate to restore muscles [6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Long Non-Coding RNA H19 Promotes Porcine Satellite Cell Differentiation by Interacting with TDP43

Zhao

et al. 2020

Genes

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Long non-coding RNAs (lncRNAs) have been implicated in fundamental and diverse biological processes, including myogenesis. However, the molecular mechanisms involved in this process remain largely unexplored. This study found that H19 affected the differentiation of porcine satellite cells (PSCs) by directly binding to the DNA/RNA-binding protein TDP43. Functional analyses showed that TDP43 knockdown decreased PSC differentiation, whereas TDP43 overexpression exerted opposite effects in vitro. Furthermore, rescue experiments demonstrated that TDP43 can rescue the decrease in PSC differentiation caused by H19 knockdown. Mechanistically, H19 may act as a scaffold to recruit TDP43 to the promoters of MYOD and thereby activate the transcription of MYOD, leading to PSC differentiation. In summary, we elucidate the molecular mechanism by which H19 and TDP43 regulate myogenesis.

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Characterization and isolation of highly purified porcine satellite cells

Cited by 86 publications

References 54 publications

Purification of Pig Muscle Stem Cells Using Magnetic-Activated Cell Sorting (MACS) Based on the Expression of Cluster of Differentiation 29 (CD29)

Purification of Pig Muscle Stem Cells Using Magnetic-Activated Cell Sorting (MACS) Based on the Expression of Cluster of Differentiation 29 (CD29)

Isolation of muscle stem cells from rat skeletal muscles

Long Non-Coding RNA H19 Promotes Porcine Satellite Cell Differentiation by Interacting with TDP43

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