Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors

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...

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Main Authors: Casey L. Roberts, Silvia S. Chen, Angela C. Murchison, Rebecca A. Ogle, Michael P. Francis, Roy C. Ogle, Patrick C. Sachs
Format: Article
Language:English
Published: Wiley 2017-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2017/1513281
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author Casey L. Roberts
Silvia S. Chen
Angela C. Murchison
Rebecca A. Ogle
Michael P. Francis
Roy C. Ogle
Patrick C. Sachs
author_facet Casey L. Roberts
Silvia S. Chen
Angela C. Murchison
Rebecca A. Ogle
Michael P. Francis
Roy C. Ogle
Patrick C. Sachs
author_sort Casey L. Roberts
collection DOAJ
description 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 PPARγ but 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.
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spelling doaj-art-e964e2c53d364dca897b493e0b6e418b2025-08-20T02:22:06ZengWileyStem Cells International1687-966X1687-96782017-01-01201710.1155/2017/15132811513281Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into OsteoprogenitorsCasey L. Roberts0Silvia S. Chen1Angela C. Murchison2Rebecca A. Ogle3Michael P. Francis4Roy C. Ogle5Patrick C. Sachs6Eastern Virginia Medical School, W. Olney Road, Norfolk, VA, USAEastern Virginia Medical School, W. Olney Road, Norfolk, VA, USAInstitute of Regenerative Medicine, LifeNet Health, Concert Drive, Virginia Beach, VA, USAInstitute of Regenerative Medicine, LifeNet Health, Concert Drive, Virginia Beach, VA, USAEastern Virginia Medical School, W. Olney Road, Norfolk, VA, USASchool of Medical Diagnostic and Translational Sciences, College of Health Sciences, Old Dominion University, Monarch Way, Norfolk, VA, USASchool of Medical Diagnostic and Translational Sciences, College of Health Sciences, Old Dominion University, Monarch Way, Norfolk, VA, USAWhile 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 PPARγ but 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.http://dx.doi.org/10.1155/2017/1513281
spellingShingle Casey L. Roberts
Silvia S. Chen
Angela C. Murchison
Rebecca A. Ogle
Michael P. Francis
Roy C. Ogle
Patrick C. Sachs
Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors
Stem Cells International
title Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors
title_full Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors
title_fullStr Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors
title_full_unstemmed Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors
title_short Preferential Lineage-Specific Differentiation of Osteoblast-Derived Induced Pluripotent Stem Cells into Osteoprogenitors
title_sort preferential lineage specific differentiation of osteoblast derived induced pluripotent stem cells into osteoprogenitors
url http://dx.doi.org/10.1155/2017/1513281
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