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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.

No MeSH data available.


Related in: MedlinePlus

Analysis of proliferation capacity, karyotype and immunophenotype of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs).(A) Mean values of cumulative population doublings. Population doublings were determined at each passage of the adherent hUCB-MSCs. Graph represents mean ± standard deviation of eight hUCB donors. (B) Representative Q-banded karyotype analysis of hUCB-MSCs in culture at passage 5 shows genetic stability of the populations (n = 3). (C) Immunophenotype of hUCB-MSCs at passage 3. Each histogram is a representative result of at least three UCB-MSC samples. Dotted lines show the IgG isotype control response and solid line curves are for the samples.
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f2: Analysis of proliferation capacity, karyotype and immunophenotype of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs).(A) Mean values of cumulative population doublings. Population doublings were determined at each passage of the adherent hUCB-MSCs. Graph represents mean ± standard deviation of eight hUCB donors. (B) Representative Q-banded karyotype analysis of hUCB-MSCs in culture at passage 5 shows genetic stability of the populations (n = 3). (C) Immunophenotype of hUCB-MSCs at passage 3. Each histogram is a representative result of at least three UCB-MSC samples. Dotted lines show the IgG isotype control response and solid line curves are for the samples.

Mentions: We obtained 127 UCB samples with a volume ranging from 10 to 158 ml. Fibroblastic adherent cells formed in 39 UCB samples (Fig. 1A), representing an isolation success rate of approximately 30% (Fig. 1B). Taking into account only samples with a minimal volume of 55 ml, the success rate improved to 57%. The results suggest that the isolation success of MSCs depends on UCB volume. Adherent cells from eight hUCB-MSCs were passaged up to 8 times, and population-doubling (PDs) and cumulative population-doublings (CPDs) were calculated. We observed an increase in CPD at each passage with an average of 19.5±0.85 at P8 (Fig. 2A). The mean PD rate of hUCB-MSCs was greater than 2.5 up to P5. However, the doubling rate decreased significantly as of P6 in which we observed a change in cell morphology. In this study, in vitro hUCB-MSC amplification up to the fifth passage did not induce alteration in the number of chromosomes or any spontaneous structural chromosomal abnormality (Fig. 2B).


Chondrogenic commitment of human umbilical cord blood-derived mesenchymal stem cells in collagen matrices for cartilage engineering
Analysis of proliferation capacity, karyotype and immunophenotype of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs).(A) Mean values of cumulative population doublings. Population doublings were determined at each passage of the adherent hUCB-MSCs. Graph represents mean ± standard deviation of eight hUCB donors. (B) Representative Q-banded karyotype analysis of hUCB-MSCs in culture at passage 5 shows genetic stability of the populations (n = 3). (C) Immunophenotype of hUCB-MSCs at passage 3. Each histogram is a representative result of at least three UCB-MSC samples. Dotted lines show the IgG isotype control response and solid line curves are for the samples.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Analysis of proliferation capacity, karyotype and immunophenotype of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs).(A) Mean values of cumulative population doublings. Population doublings were determined at each passage of the adherent hUCB-MSCs. Graph represents mean ± standard deviation of eight hUCB donors. (B) Representative Q-banded karyotype analysis of hUCB-MSCs in culture at passage 5 shows genetic stability of the populations (n = 3). (C) Immunophenotype of hUCB-MSCs at passage 3. Each histogram is a representative result of at least three UCB-MSC samples. Dotted lines show the IgG isotype control response and solid line curves are for the samples.
Mentions: We obtained 127 UCB samples with a volume ranging from 10 to 158 ml. Fibroblastic adherent cells formed in 39 UCB samples (Fig. 1A), representing an isolation success rate of approximately 30% (Fig. 1B). Taking into account only samples with a minimal volume of 55 ml, the success rate improved to 57%. The results suggest that the isolation success of MSCs depends on UCB volume. Adherent cells from eight hUCB-MSCs were passaged up to 8 times, and population-doubling (PDs) and cumulative population-doublings (CPDs) were calculated. We observed an increase in CPD at each passage with an average of 19.5±0.85 at P8 (Fig. 2A). The mean PD rate of hUCB-MSCs was greater than 2.5 up to P5. However, the doubling rate decreased significantly as of P6 in which we observed a change in cell morphology. In this study, in vitro hUCB-MSC amplification up to the fifth passage did not induce alteration in the number of chromosomes or any spontaneous structural chromosomal abnormality (Fig. 2B).

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