Epigenetic changes to human umbilical cord blood cells cultured with three proteins indicate partial reprogramming to a pluripotent state.
We have previously reported the existence of a subpopulation of cells from human umbilical cord blood capable of differentiating into oligodendrocytes, Schwann cells, bone, muscle, and endothelial cells despite their origins as CD45-positive cells. These stem cells (called FSFl cells) arise only after a period in vitro in medium containing FGF4, SCF, and Flt-3 ligand (FSFl medium) during which they express the pluripotency genes Oct4 and Nanog. The objective of this study was to determine if the novel expression of these pluripotency genes coupled with the newly acquired ability of these cells to differentiate into all three germ layers was the result of epigenetic changes to these cells after reprogramming in FSFl medium. We confirm that CD45-derived FSFl cells express Oct4 protein at levels similar to that observed among undifferentiated embryonic stem cells, during which time acetylated histones H3 and H4 display increased binding at the promoter region of Oct4. Changes to binding of acetylated histones at Oct4 when these cells are in a differentiated state (either prior to FSFl culture or after in vitro differentiation into neural cells) and when they are undifferentiated suggest that this is one way by which these cells acquire their pluripotency. While DNA hypermethylation at this gene region as well as the continued H3 and H4 acetylation at the CD45 promoter region among FSFl cells indicate this is only a partial reprogramming event, this is a significant step toward non-transgene reprogramming of somatic cells.
Wong
Christine J
CJ
Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 25 Orde Street, Toronto, Canada ON M5T 3H7.
Casper
Robert F
RF
Rogers
Ian M
IM
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D017690:Cell Transplantation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D003470:Culture Media / Q000737:chemistry; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D005312:Fetal Blood / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D019089:Stem Cell Factor / Q000235:genetics / Q000378:metabolism
D003470:Culture Media; D018398:Homeodomain Proteins; D008565:Membrane Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D019089:Stem Cell Factor; C084881:flt3 ligand protein