Isolation of muscle stem cells from rat skeletal muscles
Autor: | Amy Cortez, Francesca Boscolo Sesillo, Michelle Wong, Marianna Alperin |
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Rok vydání: | 2020 |
Předmět: |
0301 basic medicine
Hindlimb Regenerative Medicine Medical and Health Sciences 0302 clinical medicine Flow cytometry lcsh:QH301-705.5 0303 health sciences education.field_of_study Stem Cells CD29 Cell Differentiation General Medicine Skeletal Cell sorting Biological Sciences Flow Cytometry Cell biology Pelvic floor muscles CD106 medicine.anatomical_structure Muscle Female Stem Cell Research - Nonembryonic - Non-Human Stem cell 1.1 Normal biological development and functioning Population Biology Pelvic Floor Muscle Article 03 medical and health sciences Underpinning research Muscle stem cells medicine Animals education Muscle Skeletal 030304 developmental biology Cluster of differentiation Skeletal muscle Cell Biology Stem Cell Research Rats 030104 developmental biology lcsh:Biology (General) Musculoskeletal Rat PAX7 030217 neurology & neurosurgery Developmental Biology |
Zdroj: | Stem Cell Research, Vol 43, Iss, Pp-(2020) Stem cell research |
Popis: | BackgroundMuscle 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.MethodsTibialis 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.ResultsCells 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.ConclusionsCD106 is an efficient marker for reliable isolation of MuSCs from a variety of rat skeletal muscles. |
Databáze: | OpenAIRE |
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