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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 adipogenic induction of cartilage stem cells. (A) Photomicrographs of monolayer cultures following adipogenic induction of cartilage stem cells. (B) Representative examples of adipogenic induction in monolayer cultures showing 18S and LPL gene expression by RT-PCR. Control samples are unstimulated cells. Arrows indicate 200 base pair mark.
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fig9-1947603514544953: Representative samples of adipogenic induction of cartilage stem cells. (A) Photomicrographs of monolayer cultures following adipogenic induction of cartilage stem cells. (B) Representative examples of adipogenic induction in monolayer cultures showing 18S and LPL gene expression by RT-PCR. Control samples are unstimulated cells. Arrows indicate 200 base pair mark.

Mentions: After adipogenic induction Oil red-O staining was used to detect the presence of lipid vacuoles in the stem cells. All clonal cell lines from all donors had positive staining. In cultures exposed to adipogenic treatment, RT-PCR detected the expression of mRNA for lipoprotein lipase (LPL), a member of the lipase family found in adipose tissue. Control cultures had no positive staining (Fig. 9).


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 adipogenic induction of cartilage stem cells. (A) Photomicrographs of monolayer cultures following adipogenic induction of cartilage stem cells. (B) Representative examples of adipogenic induction in monolayer cultures showing 18S and LPL gene expression by RT-PCR. Control samples are unstimulated cells. Arrows indicate 200 base pair mark.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4335767&req=5

fig9-1947603514544953: Representative samples of adipogenic induction of cartilage stem cells. (A) Photomicrographs of monolayer cultures following adipogenic induction of cartilage stem cells. (B) Representative examples of adipogenic induction in monolayer cultures showing 18S and LPL gene expression by RT-PCR. Control samples are unstimulated cells. Arrows indicate 200 base pair mark.
Mentions: After adipogenic induction Oil red-O staining was used to detect the presence of lipid vacuoles in the stem cells. All clonal cell lines from all donors had positive staining. In cultures exposed to adipogenic treatment, RT-PCR detected the expression of mRNA for lipoprotein lipase (LPL), a member of the lipase family found in adipose tissue. Control cultures had no positive staining (Fig. 9).

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