Fig. 6.

Three-dimensional fibrin gel culture constructs and ASC-SF collection. (A) Fibrin gel “discs” (200 μL), plated on 24-well tissue culture plates within 7.94 mm diameter heat-stamped templates. (B) Fibrin gel “strands” (600 μL), plated between nylon tabs of FlexCell Linear TissueTrain untreated plates. Note that the discs are cultured on top of stiff tissue culture plastic while strands are cultured on top of a flexible silicone membrane. In the schematic, the long dimension is shown to illustrate the direction of constraint and dynamic stretch, however the overall geometry differs from a disc (see top view in photo). (C) ASC isolation following body-sculpting surgeries of non-smoking, non-diabetic patients below 45 years old. Stromal vascular fraction (SVF) is obtained following adipose tissue mincing and collagenase digestion, and ASCs are cultured within standard tissue culture flasks. Conditioned media (ASC-CM) is collected every 24–72 h in culture, from passages 0 to 1. (D) ASCs were sorted by flow cytometry according to staining for CD31 and CD34 cell markers, and then quantified according to percent in each subpopulation. Note the predominance of CD34⁺/CD31⁻ cells (87%) among the ASCs used for media conditioning.