Step‐Wise Chondrogenesis of Human Induced Pluripotent Stem Cells and Purification Via a Reporter Allele Generated by CRISPR‐Cas9 Genome Editing. (31st October 2018)

• Record Type: Journal Article
• Authors: Adkar, Shaunak S.; Wu, Chia‐Lung; Willard, Vincent P.; Dicks, Amanda; Ettyreddy, Adarsh; Steward, Nancy; Bhutani, Nidhi; Gersbach, Charles A.; Guilak, Farshid
• Is Part Of: Stem cells. Volume 37:Number 1(2019)
• Page Start: 65
• Page End: 76
• Publication Date: 2018-10-31
• Subjects: iPSC -- Chondrocyte -- Chondrogenesis -- CRISPR‐Cas9 -- Genome editing -- Differentiation; Cloning -- Periodicals; Clone cells -- Periodicals; Stem cells -- Periodicals; Cell Differentiation -- Periodicals; Cell Division -- Periodicals; Clone Cells -- Periodicals; Hematopoietic Stem Cells -- Periodicals; Stem Cells -- Periodicals; 571.84
• DOI: 10.1002/stem.2931
• Languages: English
• ISSNs: 1066-5099

Abstract

Abstract: 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 Abstract :