title: Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors.
abstract: While induced pluripotent stem cells (iPSCs) hold great clinical promise, one hurdle that remains is the existence of a parental germ-layer memory in reprogrammed cells leading to preferential differentiation fates. While it is problematic for generating cells vastly different from the reprogrammed cells' origins, it could be advantageous for the reliable generation of germ-layer specific cell types for future therapeutic use. Here we use human osteoblast-derived iPSCs (hOB-iPSCs) to generate induced osteoprogenitors (iOPs). Osteoblasts were successfully reprogrammed and demonstrated by endogenous upregulation of Oct4, Sox2, Nanog, TRA-1-81, TRA-16-1, SSEA3, and confirmatory hPSC Scorecard Algorithmic Assessment. The hOB-iPSCs formed embryoid bodies with cells of ectoderm and mesoderm but have low capacity to form endodermal cells. Differentiation into osteoprogenitors occurred within only 2-6 days, with a population doubling rate of less than 24 hrs; however, hOB-iPSC derived osteoprogenitors were only able to form osteogenic and chondrogenic cells but not adipogenic cells. Consistent with this, hOB-iOPs were found to have higher methylation of PPARγbut similar levels of methylation on the RUNX2 promoter. These data demonstrate that iPSCs can be generated from human osteoblasts, but variant methylation patterns affect their differentiation capacities. Therefore, epigenetic memory can be exploited for efficient generation of clinically relevant quantities of osteoprogenitor cells.
authors.affiliation: Eastern Virginia Medical School, W. Olney Road, Norfolk, VA, USA; Institute of Regenerative Medicine, LifeNet Health, Concert Drive, Virginia Beach, VA, USA.
authors.forename: Casey L
authors.initials: CL
authors.lastname: Roberts
authors.affiliation: Eastern Virginia Medical School, W. Olney Road, Norfolk, VA, USA; Institute of Regenerative Medicine, LifeNet Health, Concert Drive, Virginia Beach, VA, USA.
authors.forename: Silvia S
authors.initials: SS
authors.lastname: Chen
authors.affiliation: Institute of Regenerative Medicine, LifeNet Health, Concert Drive, Virginia Beach, VA, USA.
authors.forename: Angela C
authors.initials: AC
authors.lastname: Murchison
authors.affiliation: Institute of Regenerative Medicine, LifeNet Health, Concert Drive, Virginia Beach, VA, USA.
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Ogle
authors.affiliation: Eastern Virginia Medical School, W. Olney Road, Norfolk, VA, USA; Institute of Regenerative Medicine, LifeNet Health, Concert Drive, Virginia Beach, VA, USA.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Francis
authors.affiliation: School of Medical Diagnostic and Translational Sciences, College of Health Sciences, Old Dominion University, Monarch Way, Norfolk, VA, USA.
authors.forename: Roy C
authors.initials: RC
authors.lastname: Ogle
authors.affiliation: School of Medical Diagnostic and Translational Sciences, College of Health Sciences, Old Dominion University, Monarch Way, Norfolk, VA, USA.
authors.forename: Patrick C
authors.identifier: 0000-0002-0871-6789
authors.initials: PC
authors.lastname: Sachs