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Adipose-derived spheroid cells (Kyung Hee University)

Spheroid cells were generated from rat subcutaneous fat tissue. Cells have osteogenic and adipogenic differentiation potential. 

Rat subcutaneous tissue was carefully dissected from beneath the dermis, mechanically minced, and treated with 0.15% (w/v) collagenase type I. Collagenase was inactivated by addition of the same volume of DMEM/F12 containing  FBS (10% ). After passage through a filter of 40 -μm pore size, cells were resuspended in spheroid induction medium consisting of DMEM/F12 (3:1 ratio, v/v) supplemented with  N2 (X1),  EGF (20 ng/ml),  bFGF (20 ng/ml), and heparin (2 μg/ml ), and plated in 75 cm2 flasks.  Medium was replaced every 3 days. After 7–10 days, spheroids were harvested by centrifugation, dissociated into single cells, and resuspended in DMEM/F12 (3:1 ratio, v/v) with  FBS (10%) for monolayer culturing. When reaching high confluence, cells were harvested and replated at a ratio of 1:3 and can be cultured up to Passage 5. In order to reform spheroids, cells were incubated with spheroid induction medium for 3 days and next in neurobasal medium with G5 supplements, laminin (2 μg/ml), and  heparin (2 μg/ml), for 3–4 days.

See additional Stem, Progenitor & Primary Cells for: Adipose
Adult Stem / Progenitor Cell
Rattus norvegicus
Adipose-derived spheroid cells