Valid assessment of lipid content of cultured adipocytes using oil red O

Background: Cell culture is an important tool to study adipocyte biology. Oil red O (ORO) staining is commonly used to assess total lipid content of cultured adipocytes, as a surrogate for the degree of differentiation. However, despite widespread use of this method, its validity to measure total lipid content in the linear range has never been confirmed and published. Aim: To develop and validate a protocol that measures lipid content of cultured adipocytes in a linear range. Method: Differentiated 3T3-L1 adipocytes were washed in phosphate-buffered saline, fixed in formalin, stained with ORO, and washed severa…