Adipose tissue as a living drug: Stromal vascular fraction and adipose tissue-derived stem cells in regenerative medicine

Table 3 Summary of protocols used in different studies for enzymatic and mechanical isolation of stromal vascular fraction

Ref.	Protocols
	Enzymatic isolation of SVF
Tiryaki et al[28]	Digestive agent	Collagenase NB6, 0.1 U/mL, 1:1 (enzyme:fat, v/v)
	Incubation	37 °C for 45 minutes, gentle agitation
	Centrifugation	300 × g for 5 minutes
Agostini et al[29]	Digestive agent	Collagenase NB6 (SERVA, GMP grade), 0.15 U/mL
	Incubation	37 °C for 60-70 minutes
	Centrifugation	400 × g, for 10 minutes at 4 °C
Winnier et al[30]	Digestive agent	Matrase reagent (transpose RT kit)
	Incubation	Kit-specified
	Centrifugation	Closed system separation/filtration per kit protocol
	Mechanical isolation of SVF
Tiryaki et al[28]	Digestive agent	Pass lipoaspirate through sequential blade grids (1000 μm, 750 μm, 500 μm). Ca/Mg balanced buffer, fat:buffer, 1:3
	Incubation	10 minutes shaking after buffer addition
	Centrifugation	2000 × g for 10 minutes
Solodeev et al[25]	Digestive agent	Run actuator-driven rotating blades to mechanically disrupt tissue with 350 mL prewarmed saline
	Incubation	< 15 minutes mechanical agitation inside a closed system
	Centrifugation	400 × g for 15 minutes
Yaylacı et al[27]	Digestive agent	Pass fat repeatedly through blade grids (2.4 mm → 1.2 mm → 0.6 mm)
	Incubation	Sequential 31 + 101 passes
	Centrifugation	400 × g for 10 minutes

SVF: Stromal vascular fraction.