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Chondrogenic induction of mesenchymal stromal/stem cells from Wharton's jelly embedded in alginate hydrogel and without added growth factor: an alternative stem cell source for cartilage tissue engineering.

Reppel L, Schiavi J, Charif N, Leger L, Yu H, Pinzano A, Henrionnet C, Stoltz JF, Bensoussan D, Huselstein C - Stem Cell Res Ther (2015)

Bottom Line: We compared the results to those obtained from standard BM-MSC.From day 14 and up to 28 days, the proportion of WJ-MSC CD73(+), CD90(+), CD105(+) and CD166(+) decreased significantly compared to monolayer marker expression.Furthermore, our work highlighted a relevant result showing that WJ-MSC expressed Runx2 and type X collagen at lower levels than BM-MSC.

View Article: PubMed Central - PubMed

Affiliation: UMR 7365 CNRS-Université de Lorraine, Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), Biopôle, 54505, Vandœuvre-lès-Nancy, France. loic.reppel@univ-lorraine.fr.

ABSTRACT

Background: Due to their intrinsic properties, stem cells are promising tools for new developments in tissue engineering and particularly for cartilage tissue regeneration. Although mesenchymal stromal/stem cells from bone marrow (BM-MSC) have long been the most used stem cell source in cartilage tissue engineering, they have certain limits. Thanks to their properties such as low immunogenicity and particularly chondrogenic differentiation potential, mesenchymal stromal/stem cells from Wharton's jelly (WJ-MSC) promise to be an interesting source of MSC for cartilage tissue engineering.

Methods: In this study, we propose to evaluate chondrogenic potential of WJ-MSC embedded in alginate/hyaluronic acid hydrogel over 28 days. Hydrogels were constructed by the original spraying method. Our main objective was to evaluate chondrogenic differentiation of WJ-MSC on three-dimensional scaffolds, without adding growth factors, at transcript and protein levels. We compared the results to those obtained from standard BM-MSC.

Results: After 3 days of culture, WJ-MSC seemed to be adapted to their new three-dimensional environment without any detectable damage. From day 14 and up to 28 days, the proportion of WJ-MSC CD73(+), CD90(+), CD105(+) and CD166(+) decreased significantly compared to monolayer marker expression. Moreover, WJ-MSC and BM-MSC showed different phenotype profiles. After 28 days of scaffold culture, our results showed strong upregulation of cartilage-specific transcript expression. WJ-MSC exhibited greater type II collagen synthesis than BM-MSC at both transcript and protein levels. Furthermore, our work highlighted a relevant result showing that WJ-MSC expressed Runx2 and type X collagen at lower levels than BM-MSC.

Conclusions: Once seeded in the hydrogel scaffold, WJ-MSC and BM-MSC have different profiles of chondrogenic differentiation at both the phenotypic level and matrix synthesis. After 4 weeks, WJ-MSC, embedded in a three-dimensional environment, were able to adapt to their environment and express specific cartilage-related genes and matrix proteins. Today, WJ-MSC represent a real alternative source of stem cells for cartilage tissue engineering.

No MeSH data available.


Related in: MedlinePlus

Relative expression of specific cartilage-related genes evaluated by quantitative RT-PCR during 28 days of chondrogenic induction. All results are expressed as mean ± standard error of the mean (n ≥ 3). *p < 0.05, **p < 0.01 and ***p < 0.001, day x vs day 3 for a same cell source. ###p < 0.001, WJ-MSC vs BM-MSC for a same culture time. BM-MSC bone marrow-derived mesenchymal stromal/stem cells, COMP cartilage oligomeric matrix protein, COL collagen, WJ-MSC Wharton’s jelly-derived mesenchymal stromal/stem cells
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Fig4: Relative expression of specific cartilage-related genes evaluated by quantitative RT-PCR during 28 days of chondrogenic induction. All results are expressed as mean ± standard error of the mean (n ≥ 3). *p < 0.05, **p < 0.01 and ***p < 0.001, day x vs day 3 for a same cell source. ###p < 0.001, WJ-MSC vs BM-MSC for a same culture time. BM-MSC bone marrow-derived mesenchymal stromal/stem cells, COMP cartilage oligomeric matrix protein, COL collagen, WJ-MSC Wharton’s jelly-derived mesenchymal stromal/stem cells

Mentions: Relative expression of specific cartilage-related genes was evaluated by quantitative RT-PCR during chondrogenic differentiation. Relative expression of other mesodermic lineage markers such as Runx2 or PPARγ was also reported (Fig. 4). While aggrecan and type IIa collagen expression seemed to increase during 3D culture of WJ-MSC, Sox9, COMP and total type II collagen expression by WJ-MSC were significantly higher compared to early stages of chondrogenesis (p < 0.01). After 28 days of chondrogenic induction, type IIa and total type II collagens were significantly more expressed by WJ-MSC than by BM-MSC (p < 0.001). Conversely, COMP expression was lower in WJ-MSC compared to standard BM-MSC (p < 0.001). Hypertrophic cartilage markers, Runx2 and type X collagen, were also analyzed during chondrogenic differentiation. Our results showed that Runx2 and type X collagen were very weakly expressed by WJ-MSC throughout culture compared to BM-MSC (p < 0.001 after 28 days of chondrogenic induction). Regardless of MSC source, no expression of adipogenic transcription factor PPARγ was detected during differentiation.Fig. 4


Chondrogenic induction of mesenchymal stromal/stem cells from Wharton's jelly embedded in alginate hydrogel and without added growth factor: an alternative stem cell source for cartilage tissue engineering.

Reppel L, Schiavi J, Charif N, Leger L, Yu H, Pinzano A, Henrionnet C, Stoltz JF, Bensoussan D, Huselstein C - Stem Cell Res Ther (2015)

Relative expression of specific cartilage-related genes evaluated by quantitative RT-PCR during 28 days of chondrogenic induction. All results are expressed as mean ± standard error of the mean (n ≥ 3). *p < 0.05, **p < 0.01 and ***p < 0.001, day x vs day 3 for a same cell source. ###p < 0.001, WJ-MSC vs BM-MSC for a same culture time. BM-MSC bone marrow-derived mesenchymal stromal/stem cells, COMP cartilage oligomeric matrix protein, COL collagen, WJ-MSC Wharton’s jelly-derived mesenchymal stromal/stem cells
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4697319&req=5

Fig4: Relative expression of specific cartilage-related genes evaluated by quantitative RT-PCR during 28 days of chondrogenic induction. All results are expressed as mean ± standard error of the mean (n ≥ 3). *p < 0.05, **p < 0.01 and ***p < 0.001, day x vs day 3 for a same cell source. ###p < 0.001, WJ-MSC vs BM-MSC for a same culture time. BM-MSC bone marrow-derived mesenchymal stromal/stem cells, COMP cartilage oligomeric matrix protein, COL collagen, WJ-MSC Wharton’s jelly-derived mesenchymal stromal/stem cells
Mentions: Relative expression of specific cartilage-related genes was evaluated by quantitative RT-PCR during chondrogenic differentiation. Relative expression of other mesodermic lineage markers such as Runx2 or PPARγ was also reported (Fig. 4). While aggrecan and type IIa collagen expression seemed to increase during 3D culture of WJ-MSC, Sox9, COMP and total type II collagen expression by WJ-MSC were significantly higher compared to early stages of chondrogenesis (p < 0.01). After 28 days of chondrogenic induction, type IIa and total type II collagens were significantly more expressed by WJ-MSC than by BM-MSC (p < 0.001). Conversely, COMP expression was lower in WJ-MSC compared to standard BM-MSC (p < 0.001). Hypertrophic cartilage markers, Runx2 and type X collagen, were also analyzed during chondrogenic differentiation. Our results showed that Runx2 and type X collagen were very weakly expressed by WJ-MSC throughout culture compared to BM-MSC (p < 0.001 after 28 days of chondrogenic induction). Regardless of MSC source, no expression of adipogenic transcription factor PPARγ was detected during differentiation.Fig. 4

Bottom Line: We compared the results to those obtained from standard BM-MSC.From day 14 and up to 28 days, the proportion of WJ-MSC CD73(+), CD90(+), CD105(+) and CD166(+) decreased significantly compared to monolayer marker expression.Furthermore, our work highlighted a relevant result showing that WJ-MSC expressed Runx2 and type X collagen at lower levels than BM-MSC.

View Article: PubMed Central - PubMed

Affiliation: UMR 7365 CNRS-Université de Lorraine, Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), Biopôle, 54505, Vandœuvre-lès-Nancy, France. loic.reppel@univ-lorraine.fr.

ABSTRACT

Background: Due to their intrinsic properties, stem cells are promising tools for new developments in tissue engineering and particularly for cartilage tissue regeneration. Although mesenchymal stromal/stem cells from bone marrow (BM-MSC) have long been the most used stem cell source in cartilage tissue engineering, they have certain limits. Thanks to their properties such as low immunogenicity and particularly chondrogenic differentiation potential, mesenchymal stromal/stem cells from Wharton's jelly (WJ-MSC) promise to be an interesting source of MSC for cartilage tissue engineering.

Methods: In this study, we propose to evaluate chondrogenic potential of WJ-MSC embedded in alginate/hyaluronic acid hydrogel over 28 days. Hydrogels were constructed by the original spraying method. Our main objective was to evaluate chondrogenic differentiation of WJ-MSC on three-dimensional scaffolds, without adding growth factors, at transcript and protein levels. We compared the results to those obtained from standard BM-MSC.

Results: After 3 days of culture, WJ-MSC seemed to be adapted to their new three-dimensional environment without any detectable damage. From day 14 and up to 28 days, the proportion of WJ-MSC CD73(+), CD90(+), CD105(+) and CD166(+) decreased significantly compared to monolayer marker expression. Moreover, WJ-MSC and BM-MSC showed different phenotype profiles. After 28 days of scaffold culture, our results showed strong upregulation of cartilage-specific transcript expression. WJ-MSC exhibited greater type II collagen synthesis than BM-MSC at both transcript and protein levels. Furthermore, our work highlighted a relevant result showing that WJ-MSC expressed Runx2 and type X collagen at lower levels than BM-MSC.

Conclusions: Once seeded in the hydrogel scaffold, WJ-MSC and BM-MSC have different profiles of chondrogenic differentiation at both the phenotypic level and matrix synthesis. After 4 weeks, WJ-MSC, embedded in a three-dimensional environment, were able to adapt to their environment and express specific cartilage-related genes and matrix proteins. Today, WJ-MSC represent a real alternative source of stem cells for cartilage tissue engineering.

No MeSH data available.


Related in: MedlinePlus