DB-MSC isolation and expansion. (a) Prototype Lipogems processor (PLG-P) mounting a Lipogems device. In this closed system, the lipoaspirate is microfragmented by using mild mechanical forces without the addition of collagenase or other enzymes/additives. (b) Schematic representation of Lipogems device. The lipoaspirate (blue circle) is subjected to a cluster reduction by pushing the fat into the device through the first filter (F1). LGP-P shaking transmits a mechanical force to the stainless steel marbles contained in the device that results into a temporary fat emulsion washed by a flux of buffer. This allows the washing buffer to cross a second filter (F2) and accumulate into the drain bag (red circle) together with oil, red blood cells, and mesenchymal stromal cells (ASCs). The pellet obtained after centrifugation was seeded in 25 cm2 flask in StemMACS medium (Miltenyi Biotec, USA) (see Materials and Methods for details).