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Tendon proper- and peritenon-derived progenitor cells have unique tenogenic properties.

Mienaltowski MJ, Adams SM, Birk DE - Stem Cell Res Ther (2014)

Bottom Line: Tendon construct ultrastructure was also compared after 45 days.It also was found that peritenon-derived progenitors secrete factor(s) stimulatory to tenocytes and tendon proper progenitors.Data demonstrate that, relative to peritenon-derived progenitors, tendon proper progenitors have greater potential for forming functional tendon-like tissue.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: Multipotent progenitor populations exist within the tendon proper and peritenon of the Achilles tendon. Progenitor populations derived from the tendon proper and peritenon are enriched with distinct cell types that are distinguished by expression of markers of tendon and vascular or pericyte origins, respectively. The objective of this study was to discern the unique tenogenic properties of tendon proper- and peritenon-derived progenitors within an in vitro model. We hypothesized that progenitors from each region contribute differently to tendon formation; thus, when incorporated into a regenerative model, progenitors from each region will respond uniquely. Moreover, we hypothesized that cell populations like progenitors were capable of stimulating tenogenic differentiation, so we generated conditioned media from these cell types to analyze their stimulatory potentials.

Methods: Isolated progenitors were seeded within fibrinogen/thrombin gel-based constructs with or without supplementation with recombinant growth/differentiation factor-5 (GDF5). Early and late in culture, gene expression of differentiation markers and matrix assembly genes was analyzed. Tendon construct ultrastructure was also compared after 45 days. Moreover, conditioned media from tendon proper-derived progenitors, peritenon-derived progenitors, or tenocytes was applied to each of the three cell types to determine paracrine stimulatory effects of the factors secreted from each of the respective cell types.

Results: The cell orientation, extracellular domain and fibril organization of constructs were comparable to embryonic tendon. The tendon proper-derived progenitors produced a more tendon-like construct than the peritenon-derived progenitors. Seeded tendon proper-derived progenitors expressed greater levels of tenogenic markers and matrix assembly genes, relative to peritenon-derived progenitors. However, GDF5 supplementation improved expression of matrix assembly genes in peritenon progenitors and structurally led to increased mean fibril diameters. It also was found that peritenon-derived progenitors secrete factor(s) stimulatory to tenocytes and tendon proper progenitors.

Conclusions: Data demonstrate that, relative to peritenon-derived progenitors, tendon proper progenitors have greater potential for forming functional tendon-like tissue. Furthermore, factors secreted by peritenon-derived progenitors suggest a trophic role for this cell type as well. Thus, these findings highlight the synergistic potential of including these progenitor populations in restorative tendon engineering strategies.

No MeSH data available.


Peritenon-derived progenitor conditioned media stimulates expression of tendon differentiation markers and matrix assembly genes. Treatment of tendon proper-derived progenitors and tenocytes with peritenon-derived progenitor conditioned media (PERI) resulted in increased expression of the tendon markers Scx(A) and Tnmd(B), as well as vascular marker Emcn(C). When tendon proper-derived progenitors and tenocytes were treated with PERI conditioned media, expression of matrix assembly genes increased as a trend for Bgn(D), Dcn(E), Col12a1(F) and Col11a1(K); and significantly for Col14a1(G), Col1a1(H), Col3a1(I) and Col5a1(J). Peritenon-derived progenitors were not stimulated by tendon proper-derived progenitor or tenocyte conditioned media. (Biological replicates, n = 4 to 8; Mann–Whitney-Wilcoxon test relative to control media for each cell type for each gene assayed – P <0.01, *; P <0.05, #).
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Figure 7: Peritenon-derived progenitor conditioned media stimulates expression of tendon differentiation markers and matrix assembly genes. Treatment of tendon proper-derived progenitors and tenocytes with peritenon-derived progenitor conditioned media (PERI) resulted in increased expression of the tendon markers Scx(A) and Tnmd(B), as well as vascular marker Emcn(C). When tendon proper-derived progenitors and tenocytes were treated with PERI conditioned media, expression of matrix assembly genes increased as a trend for Bgn(D), Dcn(E), Col12a1(F) and Col11a1(K); and significantly for Col14a1(G), Col1a1(H), Col3a1(I) and Col5a1(J). Peritenon-derived progenitors were not stimulated by tendon proper-derived progenitor or tenocyte conditioned media. (Biological replicates, n = 4 to 8; Mann–Whitney-Wilcoxon test relative to control media for each cell type for each gene assayed – P <0.01, *; P <0.05, #).

Mentions: During repair, several cell types present in the lesion offer the opportunity for crosstalk to occur among cells, thus affecting re-establishment of tendon form and function. To test the hypotheses that different progenitor populations present in healing tissue are capable of cross talk in the regenerative response, an in vitro monolayer culture model was utilized. Specifically, a conditioned media experiment was done to analyze interactions via paracrine stimulatory products. Gene expression analyses were used to discern the effects of the cell type-specific secretory products of peritenon-derived progenitors, tendon proper-derived progenitors, and tenocytes, resulting in nine cell type and media combinations - cell type (conditioned media): TP (control), TP (PERI), TP (Tn), PERI (control), PERI (TP), PERI (Tn), Tn (control), Tn (TP), Tn (PERI). Treatment of tendon proper-derived progenitors and tenocytes with peritenon-derived progenitor conditioned media resulted in increased expression of the tendon markers Scx (Figure 7A) and Tnmd (Figure 7B), as well as vascular marker Emcn (Figure 7C). When tendon proper-derived progenitors and tenocytes were treated with peritenon-derived progenitor conditioned media (PERI), expression of matrix assembly genes increased as a trend for Bgn (Figure 7D), Dcn (Figure 7E) and Col12a1 (Figure 7F). Significant increases in expression were demonstrated for Col14a1 (Figure 7G), Col1a1 (Figure 7H), Col3a1 (Figure 7I), Col5a1 (Figure 7J) and Col11a1 (Figure 7K) in tendon proper-derived progenitors and tenocytes when treated with peritenon-derived progenitor conditioned media. However, peritenon-derived progenitors were not stimulated by tendon proper-derived progenitor or tenocyte conditioned media.


Tendon proper- and peritenon-derived progenitor cells have unique tenogenic properties.

Mienaltowski MJ, Adams SM, Birk DE - Stem Cell Res Ther (2014)

Peritenon-derived progenitor conditioned media stimulates expression of tendon differentiation markers and matrix assembly genes. Treatment of tendon proper-derived progenitors and tenocytes with peritenon-derived progenitor conditioned media (PERI) resulted in increased expression of the tendon markers Scx(A) and Tnmd(B), as well as vascular marker Emcn(C). When tendon proper-derived progenitors and tenocytes were treated with PERI conditioned media, expression of matrix assembly genes increased as a trend for Bgn(D), Dcn(E), Col12a1(F) and Col11a1(K); and significantly for Col14a1(G), Col1a1(H), Col3a1(I) and Col5a1(J). Peritenon-derived progenitors were not stimulated by tendon proper-derived progenitor or tenocyte conditioned media. (Biological replicates, n = 4 to 8; Mann–Whitney-Wilcoxon test relative to control media for each cell type for each gene assayed – P <0.01, *; P <0.05, #).
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Related In: Results  -  Collection

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Figure 7: Peritenon-derived progenitor conditioned media stimulates expression of tendon differentiation markers and matrix assembly genes. Treatment of tendon proper-derived progenitors and tenocytes with peritenon-derived progenitor conditioned media (PERI) resulted in increased expression of the tendon markers Scx(A) and Tnmd(B), as well as vascular marker Emcn(C). When tendon proper-derived progenitors and tenocytes were treated with PERI conditioned media, expression of matrix assembly genes increased as a trend for Bgn(D), Dcn(E), Col12a1(F) and Col11a1(K); and significantly for Col14a1(G), Col1a1(H), Col3a1(I) and Col5a1(J). Peritenon-derived progenitors were not stimulated by tendon proper-derived progenitor or tenocyte conditioned media. (Biological replicates, n = 4 to 8; Mann–Whitney-Wilcoxon test relative to control media for each cell type for each gene assayed – P <0.01, *; P <0.05, #).
Mentions: During repair, several cell types present in the lesion offer the opportunity for crosstalk to occur among cells, thus affecting re-establishment of tendon form and function. To test the hypotheses that different progenitor populations present in healing tissue are capable of cross talk in the regenerative response, an in vitro monolayer culture model was utilized. Specifically, a conditioned media experiment was done to analyze interactions via paracrine stimulatory products. Gene expression analyses were used to discern the effects of the cell type-specific secretory products of peritenon-derived progenitors, tendon proper-derived progenitors, and tenocytes, resulting in nine cell type and media combinations - cell type (conditioned media): TP (control), TP (PERI), TP (Tn), PERI (control), PERI (TP), PERI (Tn), Tn (control), Tn (TP), Tn (PERI). Treatment of tendon proper-derived progenitors and tenocytes with peritenon-derived progenitor conditioned media resulted in increased expression of the tendon markers Scx (Figure 7A) and Tnmd (Figure 7B), as well as vascular marker Emcn (Figure 7C). When tendon proper-derived progenitors and tenocytes were treated with peritenon-derived progenitor conditioned media (PERI), expression of matrix assembly genes increased as a trend for Bgn (Figure 7D), Dcn (Figure 7E) and Col12a1 (Figure 7F). Significant increases in expression were demonstrated for Col14a1 (Figure 7G), Col1a1 (Figure 7H), Col3a1 (Figure 7I), Col5a1 (Figure 7J) and Col11a1 (Figure 7K) in tendon proper-derived progenitors and tenocytes when treated with peritenon-derived progenitor conditioned media. However, peritenon-derived progenitors were not stimulated by tendon proper-derived progenitor or tenocyte conditioned media.

Bottom Line: Tendon construct ultrastructure was also compared after 45 days.It also was found that peritenon-derived progenitors secrete factor(s) stimulatory to tenocytes and tendon proper progenitors.Data demonstrate that, relative to peritenon-derived progenitors, tendon proper progenitors have greater potential for forming functional tendon-like tissue.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Introduction: Multipotent progenitor populations exist within the tendon proper and peritenon of the Achilles tendon. Progenitor populations derived from the tendon proper and peritenon are enriched with distinct cell types that are distinguished by expression of markers of tendon and vascular or pericyte origins, respectively. The objective of this study was to discern the unique tenogenic properties of tendon proper- and peritenon-derived progenitors within an in vitro model. We hypothesized that progenitors from each region contribute differently to tendon formation; thus, when incorporated into a regenerative model, progenitors from each region will respond uniquely. Moreover, we hypothesized that cell populations like progenitors were capable of stimulating tenogenic differentiation, so we generated conditioned media from these cell types to analyze their stimulatory potentials.

Methods: Isolated progenitors were seeded within fibrinogen/thrombin gel-based constructs with or without supplementation with recombinant growth/differentiation factor-5 (GDF5). Early and late in culture, gene expression of differentiation markers and matrix assembly genes was analyzed. Tendon construct ultrastructure was also compared after 45 days. Moreover, conditioned media from tendon proper-derived progenitors, peritenon-derived progenitors, or tenocytes was applied to each of the three cell types to determine paracrine stimulatory effects of the factors secreted from each of the respective cell types.

Results: The cell orientation, extracellular domain and fibril organization of constructs were comparable to embryonic tendon. The tendon proper-derived progenitors produced a more tendon-like construct than the peritenon-derived progenitors. Seeded tendon proper-derived progenitors expressed greater levels of tenogenic markers and matrix assembly genes, relative to peritenon-derived progenitors. However, GDF5 supplementation improved expression of matrix assembly genes in peritenon progenitors and structurally led to increased mean fibril diameters. It also was found that peritenon-derived progenitors secrete factor(s) stimulatory to tenocytes and tendon proper progenitors.

Conclusions: Data demonstrate that, relative to peritenon-derived progenitors, tendon proper progenitors have greater potential for forming functional tendon-like tissue. Furthermore, factors secreted by peritenon-derived progenitors suggest a trophic role for this cell type as well. Thus, these findings highlight the synergistic potential of including these progenitor populations in restorative tendon engineering strategies.

No MeSH data available.