Primary human muscle satellite cell culture: variations of cell yield, proliferation and differentiation rates according to age and sex of donors, site of muscle biopsy, and delay before processing.
The present study was performed to determine the influence on human satellite cell yield, proliferation, and differentiation rates of: 1) sex and age of donors; 2) site of the muscle biopsy; and 3) delay before processing of the muscle biopsy sample. We used a standardized primary muscle cell culture procedure on 206 normal muscle samples obtained from different muscle groups of patients aged from 20 to 88 years, at time of orthopedic surgery. Sex of donors did not influence muscle culture parameters. In contrast, aging tended to affect muscle cell yield (age group 50-59 years vs 70-79 years, P < 0.08), but not myogenic cell abilities to proliferate and to fuse into myotubes. The anatomic origin of muscle samples used for culture appeared to influence culture parameters. In contrast with other tested muscles, the tensor fasciae muscle gave both a good cell yield (174 +/- 25 10(3) cells per gram) and homogeneous proliferation and differentiation rates. Storage of the muscle sample at 4 degrees C in transport medium was associated with a very high cell yield when processing was done in early hours after biopsy (277 +/- 50 10(3) cells/g), a high and stable cell yield when processing was done from day 1 to day 3 after biopsy (185 +/- 15 10(3) cells/g), and a poor cell yield when processing was done after day 4 (111 +/- 13 10(3) cells/g). Storage of muscle biopsy samples at 4 degrees C for 1 to 4 days was associated with good proliferation and fusion rates. In conclusion, these data validate a convenient procedure of primary human muscle cell culture, using tensor fasciae muscle biopsy, which is easily done at time of orthopedic surgery, obtained from men and women of all ages (if possible less than 70 years to obtain good cell yield), and allowing of 1-3 days of storage before processing that may compensate uncertainty of the exact time of availability of muscle samples for the scientist.
Bonavaud
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Groupe d'Etudes et de Recherches sur le Muscle et le Nerf (GERMEN: ER 269+ 315), Faculté de Médecine de Créteil, Université Paris XII-Val-de-Marne.
Thibert
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Gherardi
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RK
Barlovatz-Meimon
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D000328:Adult; D000367:Age Factors; D000368:Aged; D000369:Aged, 80 and over; D002452:Cell Count; D002454:Cell Differentiation; D002455:Cell Division; D002459:Cell Fusion; D002478:Cells, Cultured; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009132:Muscles / Q000166:cytology*; D012737:Sex Factors; D013997:Time Factors