Abstract for presentation at Australia and New Zealand Society of Nephrology Annual Scientific Conference

Renal differentiation of human embryonic stem cells in serum-free culture conditions

  • Mr Navin Mahadevan, Monash University, Recipient of a 2005 US Fulbright Scholarship through the Australian-American Fulbright Commission, Australia
  • Ms Adelia Lin, Monash Institute of Medical Research, Monash University, and the Australian Stem Cell Centre, Australia
  • Mr George Zhou, Monash Institute of Medical Research, Monash University, and the Australian Stem Cell Centre, Australia
  • Dr Martin Pera, Monash Institute of Medical Research, Monash University, and the Australian Stem Cell Centre, Australia
  • Dr Andrew Laslett, Monash Institute of Medical Research, Monash University, and the Australian Stem Cell Centre, Australia

    • Human embryonic stem cells (hESC) can potentially differentiate into any cell in the body, and, as such, have great potential for application in regenerative medicine, but they have not yet been convincingly differentiated in vitro into renal cells. Thus, we investigated the capacity of hESC to differentiate into renal cells. Conventionally, hESC are grown in a fetal calf serum based culture system. Alternatively, hESC can be supported in media containing KnockOut serum replacer (SR), a serum-free reagent, under bulk culture conditions, in which cells are propagated in large numbers. Work in our lab has shown that reducing serum concentration in culture results in increased expression of genes important for renal development. To mimic these conditions, cells were grown under 5% SR conditions for 2 weeks and interrogated using flow cytometry for expression of cell surface markers which mark the population of cells thought to contain renal progenitor cells (Challen, et al. 2004), and for negative/low expression of GCTM-2, a marker of undifferentiated hESC. Results indicated that a higher proportion of cells grown in the 5% SR condition had differentiated along the renal lineage as compared to 20% SR conditions (levels used for maintenance of undifferentiated hESC). Quantitative RT-PCR analysis of sorted cell populations revealed decreased levels of stem cells markers (OCT-4, GDF3) and higher levels of renal genes (PAX-2, LHX1, WT1) as compared to controls. Thus, our results provide evidence for renal differentiation of hESC in a serum-free system.

    Challen et al. (2004) J. Am. Soc. Nephrol. 15, 2344-2357.

    Conference Organiser - ICMS Pty Ltd