Enzymatic Isolation and Serum-Free Culture of Human Renal Cells: Retaining Properties of Proximal Tubule Cells

Procedures for the culture of the human renal proximal tubule (HPT) cellutilizing explanted tissue have been previously reported by this laboratory (1 ). Several other investigators have also reported the isolation and culture of human renal tubule cells (2 , and references therein). Although explantation of tissue fragments remains an effective way to initiate cell cultures, cell outgrowth and the attainment of confluent cultures may take several weeks. The cell-culture methodology described in this chapter results in a high yield of confluent primary cultures in 7–10 d. The technique involves the digestion of minced cortical tissue with collagenase, followed by a filtering step to remove tissue fragments. The filtrate is centrifuged, and the cell pellet is resuspended in serum-free growth medium and dispensed onto prepared growth surfaces.