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Chondrogenic commitment of human umbilical cord blood-derived mesenchymal stem cells in collagen matrices for cartilage engineering

View Article: PubMed Central - PubMed

ABSTRACT

Umbilical cord blood (UCB) is a promising alternative source of mesenchymal stem cells (MSCs), because UCB-MSCs are abundant and harvesting them is a painless non-invasive procedure. Potential clinical applications of UCB-MSCs have been identified, but their ability for chondrogenic differentiation has not yet been fully evaluated. The aim of our work was to characterize and determine the chondrogenic differentiation potential of human UCB-MSCs (hUCB-MSCs) for cartilage tissue engineering using an approach combining 3D culture in type I/III collagen sponges and chondrogenic factors. Our results showed that UCB-MSCs have a high proliferative capacity. These cells differentiated easily into an osteoblast lineage but not into an adipocyte lineage. Furthermore, BMP-2 and TGF-β1 potentiated chondrogenic differentiation, as revealed by a strong increase in mature chondrocyte-specific mRNA (COL2A1, COL2B, ACAN) and protein (type II collagen) markers. Although growth factors increased the transcription of hypertrophic chondrocyte markers such as COL10A1 and MMP13, the cells present in the neo-tissue maintained their phenotype and did not progress to terminal differentiation and mineralization of the extracellular matrix after subcutaneous implantation in nude mice. Our study demonstrates that our culture model has efficient chondrogenic differentiation, and that hUCB-MSCs can be a reliable source for cartilage tissue engineering.

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Osteogenic and adipogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs).(A,B) After culture in osteoblastic induction medium, calcium mineralization was demonstrated by alizarin red S staining. A representative example from five adherent hUCB-MSC samples is shown (magnification ×10 and ×20). (C,D,G,H) Undifferentiated hUCB-MSCs. (E,F) After adipogenic induction and incubation in the maintenance medium, only a few cells show lipid droplets in the cytoplasm with oil red O staining. A representative example from three UCB donors is shown (magnification ×40).
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f3: Osteogenic and adipogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs).(A,B) After culture in osteoblastic induction medium, calcium mineralization was demonstrated by alizarin red S staining. A representative example from five adherent hUCB-MSC samples is shown (magnification ×10 and ×20). (C,D,G,H) Undifferentiated hUCB-MSCs. (E,F) After adipogenic induction and incubation in the maintenance medium, only a few cells show lipid droplets in the cytoplasm with oil red O staining. A representative example from three UCB donors is shown (magnification ×40).

Mentions: To investigate hUCB-MSC differentiation potential in vitro, five hUCB-MSC samples were exposed to induction media for osteogenic or adipogenic differentiation for 3 weeks. After osteogenic induction, hUCB-MSCs showed ECM mineralization detected by alizarin red S staining in all samples (Fig. 3). Regarding adipogenic differentiation, only a few cells showed oil red O-stained droplets in their cytoplasm in three samples (Fig. 3). The two remaining samples did not show any stained lipid droplets, even after applying a 5 week fortified differentiation protocol, which comprised only the adipogenic induction medium in culture. No extracellular matrix mineralization or lipid droplets were detected in negative controls.


Chondrogenic commitment of human umbilical cord blood-derived mesenchymal stem cells in collagen matrices for cartilage engineering
Osteogenic and adipogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs).(A,B) After culture in osteoblastic induction medium, calcium mineralization was demonstrated by alizarin red S staining. A representative example from five adherent hUCB-MSC samples is shown (magnification ×10 and ×20). (C,D,G,H) Undifferentiated hUCB-MSCs. (E,F) After adipogenic induction and incubation in the maintenance medium, only a few cells show lipid droplets in the cytoplasm with oil red O staining. A representative example from three UCB donors is shown (magnification ×40).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5015060&req=5

f3: Osteogenic and adipogenic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs).(A,B) After culture in osteoblastic induction medium, calcium mineralization was demonstrated by alizarin red S staining. A representative example from five adherent hUCB-MSC samples is shown (magnification ×10 and ×20). (C,D,G,H) Undifferentiated hUCB-MSCs. (E,F) After adipogenic induction and incubation in the maintenance medium, only a few cells show lipid droplets in the cytoplasm with oil red O staining. A representative example from three UCB donors is shown (magnification ×40).
Mentions: To investigate hUCB-MSC differentiation potential in vitro, five hUCB-MSC samples were exposed to induction media for osteogenic or adipogenic differentiation for 3 weeks. After osteogenic induction, hUCB-MSCs showed ECM mineralization detected by alizarin red S staining in all samples (Fig. 3). Regarding adipogenic differentiation, only a few cells showed oil red O-stained droplets in their cytoplasm in three samples (Fig. 3). The two remaining samples did not show any stained lipid droplets, even after applying a 5 week fortified differentiation protocol, which comprised only the adipogenic induction medium in culture. No extracellular matrix mineralization or lipid droplets were detected in negative controls.

View Article: PubMed Central - PubMed

ABSTRACT

Umbilical cord blood (UCB) is a promising alternative source of mesenchymal stem cells (MSCs), because UCB-MSCs are abundant and harvesting them is a painless non-invasive procedure. Potential clinical applications of UCB-MSCs have been identified, but their ability for chondrogenic differentiation has not yet been fully evaluated. The aim of our work was to characterize and determine the chondrogenic differentiation potential of human UCB-MSCs (hUCB-MSCs) for cartilage tissue engineering using an approach combining 3D culture in type I/III collagen sponges and chondrogenic factors. Our results showed that UCB-MSCs have a high proliferative capacity. These cells differentiated easily into an osteoblast lineage but not into an adipocyte lineage. Furthermore, BMP-2 and TGF-β1 potentiated chondrogenic differentiation, as revealed by a strong increase in mature chondrocyte-specific mRNA (COL2A1, COL2B, ACAN) and protein (type II collagen) markers. Although growth factors increased the transcription of hypertrophic chondrocyte markers such as COL10A1 and MMP13, the cells present in the neo-tissue maintained their phenotype and did not progress to terminal differentiation and mineralization of the extracellular matrix after subcutaneous implantation in nude mice. Our study demonstrates that our culture model has efficient chondrogenic differentiation, and that hUCB-MSCs can be a reliable source for cartilage tissue engineering.

No MeSH data available.


Related in: MedlinePlus