Isolation of muscle stem cells from rat skeletal muscles

Autor: Amy Cortez, Francesca Boscolo Sesillo, Michelle Wong, Marianna Alperin
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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