Early induction of a prechondrogenic population allows efficient generation of stable chondrocytes from human induced pluripotent stem cells

Lee, Jieun; Taylor, Sarah; Smeriglio, Piera; Lai, Janice H.; Maloney, William J.; Yang, Fan; Bhutani, Nidhi

doi:10.1096/fj.14-269720

Cited by 51 publications

(42 citation statements)

References 38 publications

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“…The high efficiency of this procedure was confirmed by the Safranin “O” staining and high expression of chondrogenic markers [16]. According to Lee and collaborators [17], hiPSCs can be efficiently differentiated into chondrocytes through EBs on fibronectin-coated plates for 14 days using a cocktail of defined GFs. However, note that this protocol requires a relatively large amount of GFs in high concentrations (up to 100 ng/ml), making this technique expensive and, therefore, diminishing its potential clinical application [17].…”

Section: Discussionmentioning

confidence: 99%

Comparison of Four Protocols to Generate Chondrocyte-Like Cells from Human Induced Pluripotent Stem Cells (hiPSCs)

Suchorska

Augustyniak

Richter

et al. 2016

Stem Cell Rev and Rep

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Stem cells (SCs) are a promising approach to regenerative medicine, with the potential to treat numerous orthopedic disorders, including osteo-degenerative diseases. The development of human-induced pluripotent stem cells (hiPSCs) has increased the potential of SCs for new treatments. However, current methods of differentiating hiPSCs into chondrocyte-like cells are suboptimal and better methods are needed. The aim of the present study was to assess four different chondrogenic differentiation protocols to identify the most efficient method of generating hiPSC-derived chondrocytes. For this study, hiPSCs were obtained from primary human dermal fibroblasts (PHDFs) and differentiated into chondrocyte-like cells using four different protocols: 1) monolayer culture with defined growth factors (GF); 2) embryoid bodies (EBs) in a chondrogenic medium with TGF-β3 cells; 3) EBs in chondrogenic medium conditioned with human chondrocytes (HC-402-05a cell line) and 4) EBs in chondrogenic medium conditioned with human chondrocytes and supplemented with TGF-β3. The cells obtained through these four protocols were evaluated and compared at the mRNA and protein levels. Although chondrogenic differentiation of hiPSCs was successfully achieved with all of these protocols, the two fastest and most cost-effective methods were the monolayer culture with GFs and the medium conditioned with human chondrocytes. Both of these methods are superior to other available techniques. The main advantage of the conditioned medium is that the technique is relatively simple and inexpensive while the directed method (i.e., monolayer culture with GFs) is faster than any protocol described to date because it is does not require additional steps such as EB formation.Electronic supplementary materialThe online version of this article (doi:10.1007/s12015-016-9708-y) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Comparison of Four Protocols to Generate Chondrocyte-Like Cells from Human Induced Pluripotent Stem Cells (hiPSCs)

Suchorska

Augustyniak

Richter

et al. 2016

Stem Cell Rev and Rep

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“…We have previously established methods to differentiate hiPSC into articular-like chondrocytes (hiChondrocytes) and characterized the chondrogenic phenotype of the hiChondrocytes in terms of gene and protein expression and their ability to engineer cartilage in vitro and in vivo [10]. We therefore hypothesized that these hiChondrocytes will mimic developmentally younger chondrocytes and may have enriched levels of CD24.…”

Section: Resultsmentioning

confidence: 99%

“…Five different batches of human induced pluripotent stem cell-derived chondrocytes (hiChondrocytes) were derived by differentiation of human induced pluripotent stem cells (hiPSC) as previously described and characterized [10]. Five different batches were utilized to ensure reproducibility.…”

Section: Methodsmentioning

confidence: 99%

CD24 enrichment protects while its loss increases susceptibility of juvenile chondrocytes towards inflammation

et al. 2016

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BackgroundDiseases associated with human cartilage, including rheumatoid arthritis (RA) and osteoarthritis (OA) have manifested age, mechanical stresses and inflammation as the leading risk factors. Although inflammatory processes are known to be upregulated upon aging, we sought to gain a molecular understanding of how aging affects the tissue-specific response to inflammation. In this report, we explored the role of cluster of differentiation 24 (CD24) in regulating differential inflammatory responses in juvenile and adult human chondrocytes.MethodsDifferential cell-surface CD24 expression was assessed in juvenile and adult chondrocytes along with human induced pluripotent stem cell (hiPSC)-derived neonatal chondrocytes through gene expression and fluorescence-activated cell sorting (FACS) analyses. Loss of function of CD24 was achieved through silencing in chondrocytes and the effects on the response to inflammatory cues were assessed through gene expression and NFκB activity.ResultsCD24 expression in chondrocytes caused a differential response to cytokine-induced inflammation, with the CD24high juvenile chondrocytes being resistant to IL-1ß treatment as compared to CD24low adult chondrocytes. CD24 protects from inflammatory response by reducing NFκB activation, as an acute loss of CD24 via silencing led to an increase in NFκB activation. Moreover, the loss of CD24 in chondrocytes subsequently increased inflammatory and catabolic gene expression both in the absence and presence of IL-1ß.ConclusionsWe have identified CD24 as a novel regulator of inflammatory response in cartilage that is altered during development and aging and could potentially be therapeutic in RA and OA.Electronic supplementary materialThe online version of this article (doi:10.1186/s13075-016-1183-y) contains supplementary material, which is available to authorized users.

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“…Recent advances in chondrogenic differentiation of human iPSCs support the establishment of disease modeling platforms on which candidate DMOADs that potentially prevent or reverse tissue degradation are tested for clinical efficacy. [101–103] Subsequent disease modeling studies might ask how cell lines with edited genetic backgrounds are differentially affected by these degradative stimuli; drugs efficacious in specific backgrounds might then be identified. For example, interrogation of GDF5 rs143383 variants on OA susceptibility can be considered an attractive proof of concept test, given the known functional role of GDF5 in gene regulation, joint development, and OA pathogenesis.…”

Section: Disease Modeling Applicationsmentioning

confidence: 99%

Genome Engineering for Personalized Arthritis Therapeutics

Adkar

Brunger

Willard

et al. 2017

Trends in Molecular Medicine

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Arthritis represents a family of complex joint pathologies responsible for the majority of musculoskeletal conditions. Nearly all diseases within this family, including osteoarthritis, rheumatoid arthritis, and juvenile idiopathic arthritis, are chronic conditions with few or no disease-modifying therapeutics available. Advances in genome engineering technology, most recently with CRISPR-Cas9, have revolutionized our ability to interrogate and validate genetic and epigenetic elements associated with chronic diseases such as arthritis. These technologies, together with cell reprogramming methods, including the use of induced pluripotent stem cells, provide a platform for human disease modeling. We summarize new evidence from genome-wide association studies and genomics that substantiates a genetic basis for arthritis pathogenesis. We also review the potential contributions of genome engineering in the development of new arthritis therapeutics.

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Early induction of a prechondrogenic population allows efficient generation of stable chondrocytes from human induced pluripotent stem cells

Cited by 51 publications

References 38 publications

Comparison of Four Protocols to Generate Chondrocyte-Like Cells from Human Induced Pluripotent Stem Cells (hiPSCs)

Comparison of Four Protocols to Generate Chondrocyte-Like Cells from Human Induced Pluripotent Stem Cells (hiPSCs)

CD24 enrichment protects while its loss increases susceptibility of juvenile chondrocytes towards inflammation

Genome Engineering for Personalized Arthritis Therapeutics

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