Table 1.
. Review of various porcine muscle tissue digestion procedures from literature
| Tissue digestion with: | Dissociation medium | Concentration (C), temperature (T), incubation time (IT) | Muscle | Age of the species | SC enrichment after dissociation | References |
|---|---|---|---|---|---|---|
| Pronase | PBS, EBSS, medium |
C 0.5–1.4 mg/ml, T 37°C, IT 40–90 min |
SM, ST, BF, LD, PM, VM | Neonatal–adult | Pre-plating, Percoll gradients or none | (Alexander et al. 2010, Alexander et al. 2012, Baquero-Perez et al. 2012, Blanton et al. 1999, Doumit and Merkel 1992, Gao et al. 2017, Hathaway et al. 1991, Holzer et al. 2005, Mesires and Doumit 2002, Wilschut et al. 2008, Yi et al. 2001, Zhu et al. 2013) |
| Pronase followed by collagenase | PBS with 1% HEPES (pronase), DMEM-HG with 5% FCS (collagenase) |
C 1–1.5 mg/ml pronase, 1.5 mg/ml collagenase XI, T 37°C, IT 60 min each |
ST | Neonatal | Frequent pre-plating | (Wilschut et al. 2010a, Wilschut et al. 2010b) |
| Trypsin | Ca2+ and Mg2+- free EBSS, PBS |
C 0.2–0.25% trypsin, T 37°C, IT 60 min |
Hind limb muscle, SM, ST, LD | Fetus, neonatal | Pre-plating, Percoll gradients |
(Hembree et al. 1991, Mau et al. 2008a, Mau et al. 2008b, Miersch et al. 2017, Will et al. 2012) |
| Collagenase | PBS with CaCl2, medium |
C 0.5 mg/ml collagenase IV or 0.2% collagenase II, T 37°C, IT 60–120 min |
SM, LD, ST, diaphragm | Neonatal–juvenile | Pre-plating | (Gao et al. 2017, Redshaw and Loughna 2012, Redshaw et al. 2010, Wang et al. 2016, Wang et al. 2012) |
| Collagenase with trypsin and DNase | PBS with 1% glucose (Ca2+ free) |
C 1.5–1.9 mg/ml collagenase II, 0.25–0.31% (trypsin), 0.1–0.01% (DNase I), T 37°C, IT 3 × 20 min |
SM, LD | Fetus, neonatal | 20% Percoll gradient | (Nissen and Oksbjerg 2009, Nissen et al. 2005, Ortenblad et al. 2003, Perruchot et al. 2012, Theil et al. 2006) |
| Collagenase with dispase | PBS + 2.5 mM CaCl2, DMEM | C 0.2–1% collagenase B or D, 1.1–2.4 U/ml dispase II, T 37°C, IT 24–90 min | ST, SM | Newborn, juvenile | Frequent pre-plating | (Ding et al. 2017, Wilschut et al. 2010a) |
SM Musculus (M.) semimembranosus, ST M. semitendinosus, BF M. biceps femoris, PM M. psoas major, LD Musculus longissimus dorsi, VM M. vastus medialis