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Presence of cartilage stem/progenitor cells in adult mice auricular perichondrium.

Kobayashi S, Takebe T, Zheng YW, Mizuno M, Yabuki Y, Maegawa J, Taniguchi H - PLoS ONE (2011)

Bottom Line: Furthermore, LRCs were successfully isolated and cultivated from auricular cartilage.Immunocytochemical analyses showed that LRCs express CD44 and integrin-α(5).Further characterization and utilization of the cell population should improve our understanding of basic cartilage biology and lead to advances in cartilage tissue engineering and novel therapeutic strategies for patients with craniofacial defects, including long-term tissue restoration.

View Article: PubMed Central - PubMed

Affiliation: Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Kanazawa-ku, Yokohama, Kanagawa, Japan.

ABSTRACT

Background: Based on evidence from several other tissues, cartilage stem/progenitor cells in the auricular cartilage presumably contribute to tissue development or homeostasis of the auricle. However, no definitive studies have identified or characterized a stem/progenitor population in mice auricle.

Methodology/principal findings: The 5-bromo-2'-deoxyuridine (BrdU) label-retaining technique was used to label dividing cells in fetal mice. Observations one year following the labeling revealed that label-retaining cells (LRCs) were present specifically in auricular perichondrium at a rate of 0.08±0.06%, but LRCs were not present in chondrium. Furthermore, LRCs were successfully isolated and cultivated from auricular cartilage. Immunocytochemical analyses showed that LRCs express CD44 and integrin-α(5). These LRCs, putative stem/progenitor cells, possess clonogenicity and chondrogenic capability in vitro.

Conclusions/significance: We have identified a population of putative cartilage stem/progenitor cells in the auricular perichondrium of mice. Further characterization and utilization of the cell population should improve our understanding of basic cartilage biology and lead to advances in cartilage tissue engineering and novel therapeutic strategies for patients with craniofacial defects, including long-term tissue restoration.

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Presence of long-term label retaining cells in auricuar perichondrium 48 weeks following birth.(A) Quantification of BrdU-label retaining cells from 0 day to 48 weeks in mice auricular cartilage (N = 6). (B) The distributions of LRCs between chondrium and perichondrium layer. 4 weeks after birth, all LRCs were detected in perichondrium, but not chondrium layer. (C) Proliferation of cells in auricular cartilage. Cells in both layers extensively proliferate till 2 weeks post birth. Graph shows time course-dependent changes of Ki67-positive cell index (KI) in perichondrium and chondrium for 48 weeks after birth. ;KI of Perichondrium, ; KI of Chondrium.
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pone-0026393-g003: Presence of long-term label retaining cells in auricuar perichondrium 48 weeks following birth.(A) Quantification of BrdU-label retaining cells from 0 day to 48 weeks in mice auricular cartilage (N = 6). (B) The distributions of LRCs between chondrium and perichondrium layer. 4 weeks after birth, all LRCs were detected in perichondrium, but not chondrium layer. (C) Proliferation of cells in auricular cartilage. Cells in both layers extensively proliferate till 2 weeks post birth. Graph shows time course-dependent changes of Ki67-positive cell index (KI) in perichondrium and chondrium for 48 weeks after birth. ;KI of Perichondrium, ; KI of Chondrium.

Mentions: Then, we treated pregnant mice with BrdU on day17 to 19 of gestation to label proliferating cells in the fetuses during auricular development. The auricles of the offspring were analyzed at multiple time points (on day 0 and day 3 and at 1, 2, 4, 24, and 48 weeks) (Figure 2A–G). To assess the efficiency of BrdU incorporation, the auricles of neonate mice were immunohistochemically labeled with monoclonal antibodies against BrdU; the majority of cells (83.6±5.4%) were labeled in neonates (N = 6) (Figure 3A). Within two weeks, the number of BrdU-positive cells, i.e., label-retaining cells (LRCs), rapidly decreased to 2.5±0.2% (N = 6). Concomitantly, localization of LRCs is restricted to the perichondrium layer (50.0%, 53.8%, 80.3% and 69.7% of BrdU positive cells, respectively)(Figure 3B). After 4 weeks, no LRCs were observed in the chondrium. However, LRCs were observed in the perichondrium even after 4 weeks (0.3±0.2% of cells) (N = 3). After 24 and 48 weeks, long-term LRCs were detected only in the perichondrium (0.1±0.05% and 0.08±0.06% of all 4,6-diamidino-2-phenylindole (DAPI)-stained cells, respectively) (N = 3, each timepoint) (Figure S2A, B, and Figure 3B). These LRCs, observed only in the perichondrium layer, seemed dormant, which is characteristic of putative stem cells.


Presence of cartilage stem/progenitor cells in adult mice auricular perichondrium.

Kobayashi S, Takebe T, Zheng YW, Mizuno M, Yabuki Y, Maegawa J, Taniguchi H - PLoS ONE (2011)

Presence of long-term label retaining cells in auricuar perichondrium 48 weeks following birth.(A) Quantification of BrdU-label retaining cells from 0 day to 48 weeks in mice auricular cartilage (N = 6). (B) The distributions of LRCs between chondrium and perichondrium layer. 4 weeks after birth, all LRCs were detected in perichondrium, but not chondrium layer. (C) Proliferation of cells in auricular cartilage. Cells in both layers extensively proliferate till 2 weeks post birth. Graph shows time course-dependent changes of Ki67-positive cell index (KI) in perichondrium and chondrium for 48 weeks after birth. ;KI of Perichondrium, ; KI of Chondrium.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198405&req=5

pone-0026393-g003: Presence of long-term label retaining cells in auricuar perichondrium 48 weeks following birth.(A) Quantification of BrdU-label retaining cells from 0 day to 48 weeks in mice auricular cartilage (N = 6). (B) The distributions of LRCs between chondrium and perichondrium layer. 4 weeks after birth, all LRCs were detected in perichondrium, but not chondrium layer. (C) Proliferation of cells in auricular cartilage. Cells in both layers extensively proliferate till 2 weeks post birth. Graph shows time course-dependent changes of Ki67-positive cell index (KI) in perichondrium and chondrium for 48 weeks after birth. ;KI of Perichondrium, ; KI of Chondrium.
Mentions: Then, we treated pregnant mice with BrdU on day17 to 19 of gestation to label proliferating cells in the fetuses during auricular development. The auricles of the offspring were analyzed at multiple time points (on day 0 and day 3 and at 1, 2, 4, 24, and 48 weeks) (Figure 2A–G). To assess the efficiency of BrdU incorporation, the auricles of neonate mice were immunohistochemically labeled with monoclonal antibodies against BrdU; the majority of cells (83.6±5.4%) were labeled in neonates (N = 6) (Figure 3A). Within two weeks, the number of BrdU-positive cells, i.e., label-retaining cells (LRCs), rapidly decreased to 2.5±0.2% (N = 6). Concomitantly, localization of LRCs is restricted to the perichondrium layer (50.0%, 53.8%, 80.3% and 69.7% of BrdU positive cells, respectively)(Figure 3B). After 4 weeks, no LRCs were observed in the chondrium. However, LRCs were observed in the perichondrium even after 4 weeks (0.3±0.2% of cells) (N = 3). After 24 and 48 weeks, long-term LRCs were detected only in the perichondrium (0.1±0.05% and 0.08±0.06% of all 4,6-diamidino-2-phenylindole (DAPI)-stained cells, respectively) (N = 3, each timepoint) (Figure S2A, B, and Figure 3B). These LRCs, observed only in the perichondrium layer, seemed dormant, which is characteristic of putative stem cells.

Bottom Line: Furthermore, LRCs were successfully isolated and cultivated from auricular cartilage.Immunocytochemical analyses showed that LRCs express CD44 and integrin-α(5).Further characterization and utilization of the cell population should improve our understanding of basic cartilage biology and lead to advances in cartilage tissue engineering and novel therapeutic strategies for patients with craniofacial defects, including long-term tissue restoration.

View Article: PubMed Central - PubMed

Affiliation: Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Kanazawa-ku, Yokohama, Kanagawa, Japan.

ABSTRACT

Background: Based on evidence from several other tissues, cartilage stem/progenitor cells in the auricular cartilage presumably contribute to tissue development or homeostasis of the auricle. However, no definitive studies have identified or characterized a stem/progenitor population in mice auricle.

Methodology/principal findings: The 5-bromo-2'-deoxyuridine (BrdU) label-retaining technique was used to label dividing cells in fetal mice. Observations one year following the labeling revealed that label-retaining cells (LRCs) were present specifically in auricular perichondrium at a rate of 0.08±0.06%, but LRCs were not present in chondrium. Furthermore, LRCs were successfully isolated and cultivated from auricular cartilage. Immunocytochemical analyses showed that LRCs express CD44 and integrin-α(5). These LRCs, putative stem/progenitor cells, possess clonogenicity and chondrogenic capability in vitro.

Conclusions/significance: We have identified a population of putative cartilage stem/progenitor cells in the auricular perichondrium of mice. Further characterization and utilization of the cell population should improve our understanding of basic cartilage biology and lead to advances in cartilage tissue engineering and novel therapeutic strategies for patients with craniofacial defects, including long-term tissue restoration.

Show MeSH
Related in: MedlinePlus