Application of floating cells for improved harvest in human chondrocyte culture

Cell culture medium, which must be discarded during medium change, may contain many cells that do not attach to culture plates. In the present study, we focused on these floating cells and attempted to determine their usefulness for cartilage regeneration. We counted the number of floating cells discarded during medium change and compared the proliferation and differentiation between floating cells and their adherent counterparts. Chondrocyte monolayer culture at a density of 5 × 103 cells/cm(2) produced viable floating cells at a rate of 2.7-3.2 × 10(3) cells/cm(2) per primary culture. When only the floating cells from one dish were harvested and replated in another dish, the number of cells was 2.8 × 10(4) cells/cm(2) (approximately half confluency) on culture day 7. The number of cells was half of that obtained by culturing only adherent cells (5 × 10(4) cells/cm(2)). The floating and adherent cells showed similar proliferation and differentiation properties. The recovery of floating cells from the culture medium could provide an approximately 1.5-fold increase in cell number over conventional monolayer culture. Thus, the collection of floating cells may be regarded as a simple, easy, and reliable method to increase the cell harvest for chondrocytes.