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Evidence of a Viable Pool of Stem Cells within Human Osteoarthritic Cartilage.

Nelson L, McCarthy HE, Fairclough J, Williams R, Archer CW - Cartilage (2014)

Bottom Line: Colony forming cells were successfully isolated from osteoarthritic cartilage and extensively expanded in monolayer culture.Colony forming efficiencies were consistently below 0.1%.Clonally derived cell lines also demonstrated in vitro multilineage potential via successful differentiation into chondrogenic, osteogenic, and adipogenic lineages.

View Article: PubMed Central - PubMed

Affiliation: College of Medicine, Swansea University, Swansea, UK.

ABSTRACT

Objectives: Osteoarthritis (OA) is a debilitating disease affecting more than 4 million people in the United Kingdom. Despite its prevalence, there is no successful cell-based therapy currently used to treat patients whose cartilage is deemed irrecoverable. The present study aimed to isolate stem cells from tibial plateaux cartilage obtained from patients who underwent total knee replacements for OA and investigate their stem cell characteristics.

Design: Clonally derived cell lines were selected using a differential adhesion assay to fibronectin and expanded in monolayer culture. Colony forming efficiencies and growth kinetics were investigated. The potential for tri-lineage differentiation into chondrogenic, osteogenic, and adipogenic phenotypes were analyzed using histological stains, immunocytochemistry, and reverse transcriptase polymerase chain reaction.

Results: Colony forming cells were successfully isolated from osteoarthritic cartilage and extensively expanded in monolayer culture. Colony forming efficiencies were consistently below 0.1%. Clonal cell lines were expanded beyond 40 population doublings but disparities were observed in the number of population doublings per day. Clonally derived cell lines also demonstrated in vitro multilineage potential via successful differentiation into chondrogenic, osteogenic, and adipogenic lineages. However, variation in the degree of differentiation was observed between these clonal cell lines.

Conclusions: A viable pool of cells with stem cell characteristics have been identified within human osteoarthritic cartilage. Variation in the degree of differentiation suggests the possibility of further subpopulations of cells. The identification of this stem cell population highlights the reparative potential of these cells in osteoarthritic cartilage, which could be further exploited to aid the field of regenerative medicine.

No MeSH data available.


Related in: MedlinePlus

Representative samples of osteogenic induction of cartilage stem cells. (A and B) Osteogenic pellets stained using von Kossa for mineral deposits. (A) An abundance of mineral deposits identified throughout the osoteogenic pellet. (B) Sparse deposits within the osteogenic pellet. (C) Control pellet. A-C Scale bars = 50 µm (insert = 100 µm). (D) Representative examples of osteogenic induction in pellets showing 18S and osteonectin gene expression by RT-PCR. Arrows indicate 200 base pair mark.
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fig8-1947603514544953: Representative samples of osteogenic induction of cartilage stem cells. (A and B) Osteogenic pellets stained using von Kossa for mineral deposits. (A) An abundance of mineral deposits identified throughout the osoteogenic pellet. (B) Sparse deposits within the osteogenic pellet. (C) Control pellet. A-C Scale bars = 50 µm (insert = 100 µm). (D) Representative examples of osteogenic induction in pellets showing 18S and osteonectin gene expression by RT-PCR. Arrows indicate 200 base pair mark.

Mentions: Osteogenic induction of OA cartilage stem cells was also carried out in pelleted clonal cell lines. The pellets resembled those cultured in chondrogenic conditions in size and shape. Fixed, paraffin-embedded pellets were sectioned and stained using the von Kossa technique to detect deposits of calcium, indicative of a mineral-rich matrix. Different levels of such deposits were apparent between different clones after the 3-week incubation time. The varying levels of mineral deposits did not correspond to the varying levels of chondrogenic markers, in that it was not possible to identify pellets that were consistently poor throughout the different lineages. No positive stain was observed in any of the corresponding controls (Fig. 8A-C). By RT-PCR, positive gene expression of the osteogenic marker osteonectin was found in the osteogenically induced clones (Fig. 8D).


Evidence of a Viable Pool of Stem Cells within Human Osteoarthritic Cartilage.

Nelson L, McCarthy HE, Fairclough J, Williams R, Archer CW - Cartilage (2014)

Representative samples of osteogenic induction of cartilage stem cells. (A and B) Osteogenic pellets stained using von Kossa for mineral deposits. (A) An abundance of mineral deposits identified throughout the osoteogenic pellet. (B) Sparse deposits within the osteogenic pellet. (C) Control pellet. A-C Scale bars = 50 µm (insert = 100 µm). (D) Representative examples of osteogenic induction in pellets showing 18S and osteonectin gene expression by RT-PCR. Arrows indicate 200 base pair mark.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4335767&req=5

fig8-1947603514544953: Representative samples of osteogenic induction of cartilage stem cells. (A and B) Osteogenic pellets stained using von Kossa for mineral deposits. (A) An abundance of mineral deposits identified throughout the osoteogenic pellet. (B) Sparse deposits within the osteogenic pellet. (C) Control pellet. A-C Scale bars = 50 µm (insert = 100 µm). (D) Representative examples of osteogenic induction in pellets showing 18S and osteonectin gene expression by RT-PCR. Arrows indicate 200 base pair mark.
Mentions: Osteogenic induction of OA cartilage stem cells was also carried out in pelleted clonal cell lines. The pellets resembled those cultured in chondrogenic conditions in size and shape. Fixed, paraffin-embedded pellets were sectioned and stained using the von Kossa technique to detect deposits of calcium, indicative of a mineral-rich matrix. Different levels of such deposits were apparent between different clones after the 3-week incubation time. The varying levels of mineral deposits did not correspond to the varying levels of chondrogenic markers, in that it was not possible to identify pellets that were consistently poor throughout the different lineages. No positive stain was observed in any of the corresponding controls (Fig. 8A-C). By RT-PCR, positive gene expression of the osteogenic marker osteonectin was found in the osteogenically induced clones (Fig. 8D).

Bottom Line: Colony forming cells were successfully isolated from osteoarthritic cartilage and extensively expanded in monolayer culture.Colony forming efficiencies were consistently below 0.1%.Clonally derived cell lines also demonstrated in vitro multilineage potential via successful differentiation into chondrogenic, osteogenic, and adipogenic lineages.

View Article: PubMed Central - PubMed

Affiliation: College of Medicine, Swansea University, Swansea, UK.

ABSTRACT

Objectives: Osteoarthritis (OA) is a debilitating disease affecting more than 4 million people in the United Kingdom. Despite its prevalence, there is no successful cell-based therapy currently used to treat patients whose cartilage is deemed irrecoverable. The present study aimed to isolate stem cells from tibial plateaux cartilage obtained from patients who underwent total knee replacements for OA and investigate their stem cell characteristics.

Design: Clonally derived cell lines were selected using a differential adhesion assay to fibronectin and expanded in monolayer culture. Colony forming efficiencies and growth kinetics were investigated. The potential for tri-lineage differentiation into chondrogenic, osteogenic, and adipogenic phenotypes were analyzed using histological stains, immunocytochemistry, and reverse transcriptase polymerase chain reaction.

Results: Colony forming cells were successfully isolated from osteoarthritic cartilage and extensively expanded in monolayer culture. Colony forming efficiencies were consistently below 0.1%. Clonal cell lines were expanded beyond 40 population doublings but disparities were observed in the number of population doublings per day. Clonally derived cell lines also demonstrated in vitro multilineage potential via successful differentiation into chondrogenic, osteogenic, and adipogenic lineages. However, variation in the degree of differentiation was observed between these clonal cell lines.

Conclusions: A viable pool of cells with stem cell characteristics have been identified within human osteoarthritic cartilage. Variation in the degree of differentiation suggests the possibility of further subpopulations of cells. The identification of this stem cell population highlights the reparative potential of these cells in osteoarthritic cartilage, which could be further exploited to aid the field of regenerative medicine.

No MeSH data available.


Related in: MedlinePlus