Isolation and chondroinduction of a dermis‐isolated, aggrecan‐sensitive subpopulation with high chondrogenic potential

Abstract

Objective. To develop a process that yields tissueengineered articular cartilage constructs from skinderived cells.Methods. Dermis-isolated, aggrecan-sensitive (DIAS) cells were isolated using a modified rapid adherence process. The chondrogenic potential was measured by quantitative reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistochemistry. Filamentous actin (Factin) and vinculin organization was detected using fluorescence microscopy.Results. The rapid adherence process led to a selection of DIAS cells, <10% of the entire population. DIAS cells displayed greater chondroinduction potential, as evidenced by the formation of large numbers of chondrocytic nodules on aggrecan-coated surfaces. In addition, these cells showed higher gene expression and protein production in terms of chondrocytic markers when compared with unpurified dermis cells. Similar patterns of F-actin and vinculin organization were observed between DIAS cells and chondrocytes. Threedimensional constructs from chondroinduced DIAS cells produced greater amounts of cartilage matrix than constructs from the rest of the dermis populations.Conclusion. These findings show a series of steps that work together to form tissue-engineered articular cartilage constructs using DIAS cells. Since skin presents a minimally invasive, relatively abundant cell source for tissue engineering, this study offers evidence of an efficient and stable technique to form cartilage constructs for future cartilage regeneration with autologous cells from skin.

Cartilage Tissue Engineering Using Dermis Isolated Adult Stem Cells: The Use of Hypoxia during Expansion versus Chondrogenic Differentiation

“…Our group has found that adult dermal fibroblasts are capable of chondrogenic differentiation when cultured on the aggrecan, the main proteoglycan found in cartilage [13]. We showed that a subpopulation with high chondrogenic potential, termed dermis-isolated adult stem (DIAS) cells, could be selected through rapid adherence to tissue culture polystyrene surfaces [18], [19]. We also demonstrated the ability of DIAS cells to not only produce cartilage-like extracellular matrix (ECM), but also form tissue engineered constructs with robust mechanical properties [16].…”

Enrichment and Characterization of Human Dermal Stem/Progenitor Cells by Intracellular Granularity

“…Adult stem/progenitor cells are also known to exist in the dermis [13][14][15][16][17][18][19][20]. These cells can be characterized into various cell types, including those that differentiate into adipocytes, chondrocytes, smooth muscle cells, and osteoblasts, and those that differentiate into nonmesodermal cells, including neural cells, hepatocytes, and pancreatic cells [13][14][15][16][17][18][19][20]43,44].…”

“…Adult stem cells exist in multiple tissues of the body, including the bone marrow [2][3][4][5], brain, skeletal muscle [6], adipose tissue [7,8], umbilical cord [9], and placenta [10]. The skin is the largest organ in the body, and its stem cells have been identified in all regions of the skin, including the epidermis [11,12], dermis [13][14][15][16][17][18][19][20], and hair follicles [21,22].…”

Harnessing Biomechanics to Develop Cartilage Regeneration Strategies

“…In a subsequent study, a protocol was developed to isolate a subpopulation that displayed greater chondroinduction potential, termed DIAS cell, from whole dermis by rapid adherence. It was demonstrated that DIAS cells can be used to form cartilage neocartilage, suggesting that skin-derived stem cells can be differentiated to chondrocyte-like cells [98].…”