A novel source of cultured podocytes

PLoS One. 2013 Dec 12;8(12):e81812. doi: 10.1371/journal.pone.0081812. eCollection 2013.

Abstract

Amniotic fluid is in continuity with multiple developing organ systems, including the kidney. Committed, but still stem-like cells from these organs may thus appear in amniotic fluid. We report having established for the first time a stem-like cell population derived from human amniotic fluid and possessing characteristics of podocyte precursors. Using a method of triple positive selection we obtained a population of cells (hAKPC-P) that can be propagated in vitro for many passages without immortalization or genetic manipulation. Under specific culture conditions, these cells can be differentiated to mature podocytes. In this work we compared these cells with conditionally immortalized podocytes, the current gold standard for in vitro studies. After in vitro differentiation, both cell lines have similar expression of the major podocyte proteins, such as nephrin and type IV collagen, that are characteristic of mature functional podocytes. In addition, differentiated hAKPC-P respond to angiotensin II and the podocyte toxin, puromycin aminonucleoside, in a way typical of podocytes. In contrast to immortalized cells, hAKPC-P have a more nearly normal cell cycle regulation and a pronounced developmental pattern of specific protein expression, suggesting their suitability for studies of podocyte development for the first time in vitro. These novel progenitor cells appear to have several distinct advantages for studies of podocyte cell biology and potentially for translational therapies.

MeSH terms

  • Amniotic Fluid / cytology*
  • Amniotic Fluid / metabolism
  • Angiotensin II / pharmacology
  • Antimetabolites, Antineoplastic / pharmacology
  • Biomarkers / metabolism
  • Cell Cycle / drug effects
  • Cell Cycle / genetics*
  • Cell Differentiation
  • Cell Proliferation
  • Cell Separation
  • Cells, Cultured
  • Collagen Type IV / genetics
  • Collagen Type IV / metabolism
  • Gene Expression
  • Gene Expression Profiling
  • Humans
  • Male
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Podocytes / cytology*
  • Podocytes / drug effects
  • Podocytes / metabolism
  • Puromycin Aminonucleoside / pharmacology

Substances

  • Antimetabolites, Antineoplastic
  • Biomarkers
  • Collagen Type IV
  • Membrane Proteins
  • nephrin
  • Angiotensin II
  • Puromycin Aminonucleoside

Grant support

Research funded by: Alport Syndrome Foundation, California Institute for Regenerative Medicine Training Grant TG2 01168 and the GOFARR Foundation at Children’s Hospital Los Angeles. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.