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.

本实验室先前已报道了利用外植组织培养人肾近端小管（HPT）细胞的方法（1）。其他一些研究人员也报道了人肾小管细胞的分离和培养（2以及其中的参考文献）。尽管组织碎片的外植仍然是启动细胞培养的一种有效方法，但细胞生长以及获得融合的培养物可能需要数周时间。本章所述的细胞培养方法可在7 - 10天内获得大量融合的原代培养物。该技术包括用胶原酶消化切碎的皮质组织，随后进行过滤步骤以去除组织碎片。将滤液离心，然后将细胞沉淀重悬于无血清生长培养基中，并分配到准备好的生长表面上。