PMID- 27038912
OWN - NLM
STAT- MEDLINE
DCOM- 20160930
LR  - 20220318
IS  - 1472-6750 (Electronic)
IS  - 1472-6750 (Linking)
VI  - 16
DP  - 2016 Apr 2
TI  - Depleting extracellular vesicles from fetal bovine serum alters proliferation and 
      differentiation of skeletal muscle cells in vitro.
PG  - 32
LID - 10.1186/s12896-016-0262-0 [doi]
LID - 32
AB  - BACKGROUND: Fetal bovine serum (FBS) contains a wide range of growth factors, 
      hormones, vitamins, amino acids, fatty acids and trace elements required for cell 
      growth. It was shown that animal sera contain also extracellular vesicles (EVs) 
      with important biological properties; thus we wondered whether EVs present in FBS 
      would influence muscle cell phenotype. EVs were removed from sera by 
      ultracentrifugation (18 h). C2C12, L6 and human primary myoblasts, were grown 
      either in classical media (CM) or in EVs-depleted media. Differentiation was 
      induced by replacing the culture medium either with CM or EV-depleted media. 
      qRT-PCR of relevant genes and miRNA involved in proliferation, differentiation, 
      energy metabolism and EVs formation and secretion were performed. RESULTS: Growth 
      of myoblasts in EV-free media during proliferation produces the most unfavorable 
      situation for proper myotube formation, when considering C212 and human 
      myoblasts. Removing EVs from serum committed myoblasts to differentiate 
      precociously (induction of myogenin and decreased expression of myomiR involved 
      in myogenesis). C2C12 and human myoblasts, grown constantly in EV-depleted media 
      during proliferation and differentiation, formed less myotubes than in CM. They 
      had a reduced level of myogenin and a strong increase in myostatin expression, a 
      negative regulator of muscle cell differentiation that affects myotube size. This 
      situation was not reversed when confluent myoblasts were switched to CM for 
      differentiation. Like C2C12 and human cells, L6 formed less myotubes in 
      EVs-depleted media. However, as they do not express myostatin, L6 myotubes were 
      larger and expressed higher level of CKTM2 compared to myotubes grown in CM 
      suggesting that they had reached a higher level of differentiation. CONCLUSIONS: 
      Researchers studying the role of muscle EVs in culture conditions should consider 
      that depleting EVs from serum alters the phenotype of muscle cells. 
      Interestingly, the cross-talk between myoblasts and myotubes during myogenesis 
      (Forterre 2014, PLoS One. 2014 Jan 2;9(1):e84153) can be recapitulate by using 
      FBS-EVs as well. This implies that EVs can transfer specific signals to cells 
      from unrelated species and that part of serum EV composition is evolutionarily 
      conserved (e.g.; myomiR are detected in FBS-EVs). EVs in body fluids could have 
      an unsuspected function during embryogenesis and in regulation of cellular 
      processes such as hypertrophy and hyperplasia.
FAU - Aswad, Hala
AU  - Aswad H
AD  - CarMeN laboratory (INSERM 1060, INRA 1397, INSA), Faculte de Medecine Lyon-Sud, 
      University of Lyon, Chemin du Grand Revoyet, Oullins, 69600, France.
FAU - Jalabert, Audrey
AU  - Jalabert A
AD  - CarMeN laboratory (INSERM 1060, INRA 1397, INSA), Faculte de Medecine Lyon-Sud, 
      University of Lyon, Chemin du Grand Revoyet, Oullins, 69600, France.
FAU - Rome, Sophie
AU  - Rome S
AD  - CarMeN laboratory (INSERM 1060, INRA 1397, INSA), Faculte de Medecine Lyon-Sud, 
      University of Lyon, Chemin du Grand Revoyet, Oullins, 69600, France. 
      srome@univ-lyon1.fr.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160402
PL  - England
TA  - BMC Biotechnol
JT  - BMC biotechnology
JID - 101088663
RN  - 0 (Culture Media)
SB  - IM
MH  - Animals
MH  - Cattle
MH  - Cell Differentiation/*drug effects
MH  - Cell Line
MH  - Cell Proliferation/*drug effects
MH  - Culture Media/chemistry/*pharmacology
MH  - Extracellular Vesicles/*physiology
MH  - Humans
MH  - Muscle, Skeletal/*cytology
MH  - Serum/*chemistry
PMC - PMC4818850
OTO - NOTNLM
OT  - Bovine serum
OT  - C2C12
OT  - Extracellular vesicles
OT  - Gene expression
OT  - Human myoblasts
OT  - L6
OT  - Muscle cell
OT  - Proliferation
OT  - miRNAs
EDAT- 2016/04/04 06:00
MHDA- 2016/10/01 06:00
PMCR- 2016/04/02
CRDT- 2016/04/04 06:00
PHST- 2015/05/19 00:00 [received]
PHST- 2016/03/16 00:00 [accepted]
PHST- 2016/04/04 06:00 [entrez]
PHST- 2016/04/04 06:00 [pubmed]
PHST- 2016/10/01 06:00 [medline]
PHST- 2016/04/02 00:00 [pmc-release]
AID - 10.1186/s12896-016-0262-0 [pii]
AID - 262 [pii]
AID - 10.1186/s12896-016-0262-0 [doi]
PST - epublish
SO  - BMC Biotechnol. 2016 Apr 2;16:32. doi: 10.1186/s12896-016-0262-0.