What are the key points relating to proliferation differentiation and culturing of HWP?

PromoCell's Normal Human White Preadipocytes (HWP) are isolated from adult subcutaneous or visceral adipose tissue from different locations. The cells are frozen in our serum-free freezing medium (Cryo-SFM) at the end of passage 1 (= secondary culture). A randomly selected vial is then used for quality control; which includes determination of growth characteristics; control of morphology; and tests for differentiation capacity into mature adipocytes. The recommended seeding density of preadipocytes after thawing/trypsinization is 5;000 cells/cm²; cells should be trypsinized before reaching 90% confluence. Population doubling times are usually between 20-50 hrs (10 population doublings guaranteed). Using a 1:4 split ratio; you can perform ∼4-5 passages with the cells. Preadipocyte Growth Medium (C-27417) is used to propagate the cells. To induce differentiation of preadipocytes into adipocytes; cells are grown in PromoCell's Preadipocyte Growth Medium until they reach 100% confluency. Cells are then cultured in Preadipocyte Differentiation Medium (C-27437) for 72 h; followed by 10-14 days in Adipocyte Nutrition Medium (C-27439). During this time the cells start to accumulate fat droplets which can be visualized under the microscope. We recommend performing differentiation experiments at population doubling numbers lower than 4-5; in order to reach a high differentiation level of the culture.