Propagation of Human Embryonic Stem Cells on Human Amniotic Fluid Cells as Feeder Cells in Xeno-Free Culture Conditions.
Human embryonic stem cells (hESCs) have been routinely cultured on mouse embryonic fibroblast feederlayers with a medium containing animal materials. For clinical application of hESCs, animal-derived products from the animal feeder cells, animal substrates such as gelatin or Matrigel and animal serum are strictly to be eliminated in the culture system. In this study, we performed that SNUhES32 and H1 were cultured on human amniotic fluid cells (hAFCs) with KOSR XenoFree and a humanized substrate. All of hESCs were relatively well propagated on hAFCs feeders with xeno-free conditions and they expressed pluripotent stem cell markers, alkaline phosphatase, SSEA-4, TRA1-60, TRA1-81, Oct-4, and Nanog like hESCs cultured on STO or human foreskin fibroblast feeders. In addition, we observed the expression of nonhuman N-glycolylneuraminic acid (Neu5GC) molecules by flow cytometry, which was xenotransplantation components of contamination in hESCs cultured on animal feeder conditions, was not detected in this xeno-free condition. In conclusion, SNUhES32 and H1 could be maintained on hAFCs for humanized culture conditions, therefore, we suggested that new xenofree conditions for clinical grade hESCs culture will be useful data in future clinical studies.
Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul 03080, Korea.
Juwon
J
Jung
Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul 03080, Korea.
Jin Ah
JA
Baek
Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul 03080, Korea.
Hye Won
HW
Seol
Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul 03080, Korea; Dept. of Obstetrics & Gynecology, College of Medicine, Seoul National University, Seoul 03080, Korea.
Young Min
YM
Choi
human amniotic fluid cells; human embryonic stem cells; xeno-free culture