Step-Wise Chondrogenesis of Human Induced Pluripotent Stem Cells and Purification Via a Reporter Allele Generated by CRISPR-Cas9 Genome Editing.

Published

Journal Article

The differentiation of human induced pluripotent stem cells (hiPSCs) to prescribed cell fates enables the engineering of patient-specific tissue types, such as hyaline cartilage, for applications in regenerative medicine, disease modeling, and drug screening. In many cases, however, these differentiation approaches are poorly controlled and generate heterogeneous cell populations. Here, we demonstrate cartilaginous matrix production in three unique hiPSC lines using a robust and reproducible differentiation protocol. To purify chondroprogenitors (CPs) produced by this protocol, we engineered a COL2A1-GFP knock-in reporter hiPSC line by CRISPR-Cas9 genome editing. Purified CPs demonstrated an improved chondrogenic capacity compared with unselected populations. The ability to enrich for CPs and generate homogenous matrix without contaminating cell types will be essential for regenerative and disease modeling applications. Stem Cells 2019;37:65-76.

Full Text

Duke Authors

Cited Authors

  • Adkar, SS; Wu, C-L; Willard, VP; Dicks, A; Ettyreddy, A; Steward, N; Bhutani, N; Gersbach, CA; Guilak, F

Published Date

  • January 2019

Published In

Volume / Issue

  • 37 / 1

Start / End Page

  • 65 - 76

PubMed ID

  • 30378731

Pubmed Central ID

  • 30378731

Electronic International Standard Serial Number (EISSN)

  • 1549-4918

International Standard Serial Number (ISSN)

  • 1066-5099

Digital Object Identifier (DOI)

  • 10.1002/stem.2931

Language

  • eng