Generation of Human Epidermal Constructs on a Collagen Layer Alone

Riva, Federica; Casasco, Andrea; Nespoli, Emanuele; Cornaglia, Antonia Icaro; Casasco, Marco; Faga, Angela; Scevola, Silvia; Mazzini, Giuliano; Calligaro, Alberto

doi:10.1089/ten.2006.0329

Cited by 15 publications

(10 citation statements)

References 67 publications

(76 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is known that fibroblasts play key roles promoting keratinocyte growth and differentiation . Additionally, the presence of calcium, serum, or both, can induce keratinocyte proliferation and differentiation . These observations are in agreement with the obtained results that clearly showed the rearrangement and phenotypic variations of hKC to be dependent on the presence of hDFb and serum in the heterotypic culture established in KSFM/MEM.…”

Section: Discussionsupporting

confidence: 92%

“…[15,58] Additionally, the presence of calcium, serum, or both, can induce keratinocyte proliferation and differentiation. [59,60] These observations are in agreement with the obtained results that clearly showed the rearrangement and phenotypic variations of hKC to be dependent on the presence of hDFb and serum in the heterotypic culture established in KSFM/MEM. In fact when culturing with KSFM/MEM, proliferative Ki-67 positive hKC were detected in the basal layer.…”

Section: Discussionsupporting

confidence: 90%

See 1 more Smart Citation

Poly(hydroxybutyrate‐co‐hydroxyvalerate) Bilayer Skin Tissue Engineering Constructs with Improved Epidermal Rearrangement

Zonari

Cerqueira

Novikoff

et al. 2014

Macromolecular Bioscience

View full text Add to dashboard Cite

Bilayer skin substitutes constitute an attractive strategy towards improved skin wound healing. Therefore, solvent casting and freeze-drying methodologies are used to produce polyhydroxybutyrate-co-hydroxyvalerate (PHBV) thin nanoporous membranes and 3D porous scaffolds that are combined in bilayer structures to recreate the epidermal and dermal layers, respectively. The combination of these methodologies allow attaining a bilayer structure with a high water retention capability and adequate mechanical properties, susceptible to enzymes degradative action. Cultures established with human keratinocytes (hKC) and dermal fibroblasts (hDFb) confirm the suitability of the PHBV structures to support cell adhesion and proliferation. Nonetheless, when co-cultured under defined conditions, hKC are able to grow and rearrange in a multilayer structure with proliferative cells in the basal layer, and cells expressing a terminal differentiation marker in the upper layer. Therefore, PHBV bilayer structures demonstrate properties that favor skin cells performance, thus representing a promising strategy to improve wound healing.

show abstract

Section: Discussionsupporting

confidence: 92%

Section: Discussionsupporting

confidence: 90%

Poly(hydroxybutyrate‐co‐hydroxyvalerate) Bilayer Skin Tissue Engineering Constructs with Improved Epidermal Rearrangement

Zonari

Cerqueira

Novikoff

et al. 2014

Macromolecular Bioscience

View full text Add to dashboard Cite

show abstract

“…Furthermore, the total duration of culture, the passage number of cells and the assumptive total cell output, all suggested that the induced cells had longterm growth potential. Intriguingly, the most significant characteristic of the induced cells was their ability to regenerate a stratified skin equivalent [Riva et al, 2007]. These results demonstrated that the morphologic, phenotypic, growth and functional characteristics of the activator-induced cells were very similar to those of epidermal stem cells, i.e.…”

Section: Discussionmentioning

confidence: 67%

Differentiated Epidermal Cells Regain the Ability to Regenerate a Skin Equivalent by Increasing the Level of β-Catenin in the Cells

Zhao

Zhang

et al. 2012

Cells Tissues Organs

View full text Add to dashboard Cite

Epidermal stem cells are of major importance for skin regeneration and tissue engineering, but differentiated epidermal cells lost their proliferative capacity and are no longer able to regenerate a skin equivalent. Here, we investigated the role of β-catenin in regulating regenerative functions of differentiated epidermal cells. Lithium chloride and a highly specific glycogen synthase kinase (GSK)-3β inhibitor were applied to induce the expression of β-catenin in differentiated epidermal cells. After a 6-day induction, the large flat-shaped cells with a small nuclear-cytoplasmic ratio had changed into small round-shaped cells with a large nuclear-cytoplasmic ratio. Phenotypic assays showed a remarkably higher expression of CK19, β₁-integrin, Oct4 and Nanog in induced cells than in the control group (p < 0.01). In addition, the results of growth and functional investigations demonstrated that the induced epidermal cells exhibited a high colony-forming ability, a long-term proliferative potential and the ability to regenerate a skin equivalent, which were regarded as the most important features of epidermal stem cells. These results suggest that the activation of β-catenin favors the reversion or dedifferentiation of differentiated epidermal cells to an immature or a less differentiated state. This study may also offer a new approach to yield enough epidermal stem cells for skin regeneration and tissue engineering.

show abstract

“…Furthermore, the total days of culture, the passage number of cells, and assumptive total cell output all suggested that dedifferentiation‐derived cells, as well as epidermal stem cells, had long‐term growth potential. Intriguingly, the most significant characteristic of dedifferentiation‐derived cells was their ability to regenerate a highly stratified skin equivalents [38]. These results demonstrated that the phenotypic and functional characteristics of dedifferentiation‐derived cells cultured in vitro were similar to epidermal stem cells.…”

Section: Discussionmentioning

confidence: 93%

Dedifferentiation-derived cells exhibit phenotypic and functional characteristics of epidermal stem cells

Zhang

Chen

et al. 2009

Journal of Cellular and Molecular Medicine

View full text Add to dashboard Cite

Differentiated epidermal cells can dedifferentiate into stem cells or stem cell-like cells in vivo. In this study, we report the isolation and characterization of dedifferentiation-derived cells. Epidermal sheets eliminated of basal stem cells were transplanted onto the skin wounds in 47 nude athymic (BALB/c-nu/nu) mice. After 5 days, cells negative for CK10 but positive for CK19 and β1-integrin emerged at the wound-neighbouring side of the epidermal sheets. Furthermore, the percentages of CK19 and β1-integrin+ cells detected by flow cytometric analysis were increased after grafting (P < 0.01) and CK10+ cells in grafted sheets decreased (P < 0.01). Then we isolated these cells on the basis of rapid adhesion to type IV collagen and found that there were 4.56% adhering cells (dedifferentiation-derived cells) in the grafting group within 10 min. The in vitro phenotypic assays showed that the expressions of CK19, β1-integrin, Oct4 and Nanog in dedifferentiation-derived cells were remarkably higher than those in the control group (differentiated epidermal cells) (P < 0.01). In addition, the results of the functional investigation of dedifferentiation-derived cells demonstrated: (1) the numbers of colonies consisting of 5–10 cells and greater than 10 cells were increased 5.9-fold and 6.7-fold, respectively, as compared with that in the control (P < 0.01); (2) more cells were in S phase and G2/M phase of the cell cycle (proliferation index values were 21.02% in control group, 45.08% in group of dedifferentiation); (3) the total days of culture (28 days versus 130 days), the passage number of cells (3 passages versus 20 passages) and assumptive total cell output (1 × 105 cells versus 1 × 1012 cells) were all significantly increased and (4) dedifferentiation-derived cells, as well as epidermal stem cells, were capable of regenerating a skin equivalent, but differentiated epidermal cells could not. These results suggested that the characteristics of dedifferentiation-derived cells cultured in vitro were similar to epidermal stem cells. This study may also offer a new approach to yield epidermal stem cells for wound repair and regeneration.

show abstract

Generation of Human Epidermal Constructs on a Collagen Layer Alone

Cited by 15 publications

References 67 publications

Poly(hydroxybutyrate‐co‐hydroxyvalerate) Bilayer Skin Tissue Engineering Constructs with Improved Epidermal Rearrangement

Poly(hydroxybutyrate‐co‐hydroxyvalerate) Bilayer Skin Tissue Engineering Constructs with Improved Epidermal Rearrangement

Differentiated Epidermal Cells Regain the Ability to Regenerate a Skin Equivalent by Increasing the Level of β-Catenin in the Cells

Dedifferentiation-derived cells exhibit phenotypic and functional characteristics of epidermal stem cells

Contact Info

Product

Resources

About