E. Vinod (Vellore, IN)
Christian Medical College PhysiologyPresenter Of 1 Presentation
P238 - Preferential Isolation and Characterization of Human Chondroprogenitors from Chondrocytes Based on CD146/CD166/CD34 Surface Markers
Abstract
Purpose
Chondrocytes are used as cellular therapy for the treatment of cartilage pathologies. However, the development of fibrocartilage repair tissue remains as an obstacle. The pursuit of an alternative cell source with enhanced potential for chondrogenesis and decreased hypertrophy led to the discovery of resident cartilage stem cells called chondroprogenitors. Our recent proof-of-concept study comparing human chondrocytes and chondroprogenitors revealed significant differences in the expression of CD146, CD166, and CD34. The results suggested isolating the chondroprogenitor cells based on the aforementioned from a pool of heterogenous chondrocytes would provide cells of higher chondrogenic potential. In line with this, our goal was to compare chondrocytes, fibronectin adhesion assay-derived chondroprogenitors, CD146+CD166+CD34-, and CD146-CD166+CD34- sorted chondrocytes, to identify the population with the highest potential for chondrogenesis.
Methods and Materials
Cartilage slices from human osteoarthritic knee joints (n=3) were subjected to enzymatic digestion to obtain chondrocytes followed by fibronectin adhesion assay to isolate chondroprogenitors. Passage 2 chondrocytes were sorted to obtain CD34- and CD166+ cells, which were further sorted to obtain CD146+ and CD146- subgroups. A total of four groups were analyzed for markers of chondrogenesis and hypertrophy using RT-PCR, trilineage differentiation, and total GAG/DNA content.
Results
Chondroprogenitors and 146+ subsets displayed higher COL2A1 expression than 146- subsets and lesser MMP-13 expression than chondrocytes. Moreover, multilineage differentiation staining revealed that they displayed minimal calcification and improved glycosaminoglycan content. GAG/DNA content validated the superiority of chondroprogenitors over chondrocytes and 146- subgroups.
Conclusion
The CD146+CD34-CD166+ sorted chondrocytes possess properties akin to chondroprogenitors and their therapeutic application in cartilage repair warrants further evaluation.
Presenter Of 2 Presentations
P-10.1.1 - Comparative Analysis of Human Migratory and Fibronectin Adhesion Derived Chondroprogenitors to Assess Superiority for Cartilage Repair
P238 - Preferential Isolation and Characterization of Human Chondroprogenitors from Chondrocytes Based on CD146/CD166/CD34 Surface Markers
Abstract
Purpose
Chondrocytes are used as cellular therapy for the treatment of cartilage pathologies. However, the development of fibrocartilage repair tissue remains as an obstacle. The pursuit of an alternative cell source with enhanced potential for chondrogenesis and decreased hypertrophy led to the discovery of resident cartilage stem cells called chondroprogenitors. Our recent proof-of-concept study comparing human chondrocytes and chondroprogenitors revealed significant differences in the expression of CD146, CD166, and CD34. The results suggested isolating the chondroprogenitor cells based on the aforementioned from a pool of heterogenous chondrocytes would provide cells of higher chondrogenic potential. In line with this, our goal was to compare chondrocytes, fibronectin adhesion assay-derived chondroprogenitors, CD146+CD166+CD34-, and CD146-CD166+CD34- sorted chondrocytes, to identify the population with the highest potential for chondrogenesis.
Methods and Materials
Cartilage slices from human osteoarthritic knee joints (n=3) were subjected to enzymatic digestion to obtain chondrocytes followed by fibronectin adhesion assay to isolate chondroprogenitors. Passage 2 chondrocytes were sorted to obtain CD34- and CD166+ cells, which were further sorted to obtain CD146+ and CD146- subgroups. A total of four groups were analyzed for markers of chondrogenesis and hypertrophy using RT-PCR, trilineage differentiation, and total GAG/DNA content.
Results
Chondroprogenitors and 146+ subsets displayed higher COL2A1 expression than 146- subsets and lesser MMP-13 expression than chondrocytes. Moreover, multilineage differentiation staining revealed that they displayed minimal calcification and improved glycosaminoglycan content. GAG/DNA content validated the superiority of chondroprogenitors over chondrocytes and 146- subgroups.
Conclusion
The CD146+CD34-CD166+ sorted chondrocytes possess properties akin to chondroprogenitors and their therapeutic application in cartilage repair warrants further evaluation.