Wharton's Jelly Stem Cells: A Novel Cell Source for Oral Mucosa and Skin Epithelia Regeneration

Abstract

Perinatal stem cells such as human umbilical cord Wharton's jelly stem cells (HWJSCs) are excellent candidates for tissue engineering because of their proliferation and differentiation capabilities. However, their differentiation potential into epithelial cells at in vitro and in vivo levels has not yet been reported. In this work we have studied the capability of HWJSCs to differentiate in vitro and in vivo to oral mucosa and skin epithelial cells using a bioactive three-dimensional model that mimics the native epithelial-mesenchymal interaction. To achieve this, primary cell cultures of HWJSCs, oral mucosa, and skin fibroblasts were obtained in order to generate a three-dimensional heterotypical model of artificial oral mucosa and skin based on fibrin-agarose biomaterials. Our results showed that the cells were unable to fully differentiate to epithelial cells in vitro. Nevertheless, in vivo grafting of the bioactive three-dimensional models demonstrated that HWJSCs were able to stratify and to express typical markers of epithelial differentiation, such as cytokeratins 1, 4, 8, and 13, plakoglobin, filaggrin, and involucrin, showing specific surface patterns. Electron microscopy analysis confirmed the presence of epithelial cell-like layers and well-formed cell-cell junctions. These results suggest that HWJSCs have the potential to differentiate to oral mucosa and skin epithelial cells in vivo and could be an appropriate novel cell source for the development of human oral mucosa and skin in tissue engineering protocols.

Biomaterials for Tendon/Ligament and Skin Regeneration

“…Once the stromas jellified, human umbilical cord Wharton's jelly stem cells (HWJSCs) were seeded on top of the skin artificial stromas and cultured. It was demonstrated that this 3D bioactive scaffold can stratify and form epithelial cell-like layers and wellformed cell-cell junctions [37]. The authors also demonstrated similar strategy can be used for regeneration of oral mucosa using human Wharton's jelly stem cells (HWJSCs)-seeded fibrin-agarose-mucosal fibroblasts.…”

Ex vivo construction of a novel model of bioengineered bladder mucosa: A preliminary study

“…The structural analogy of this model with the human bladder mucosa at both the epithelial and the stromal level leads us to conclude that the bladder mucosa produced by tissue engineering is analogous to the native tissues, although immature at this stage. Based on previous tissue models generated by the research group using fibrin‐agarose scaffolds, we hypothesize that these bladder mucosa substitutes might show positive results in vivo , and possibly have therapeutic usefulness . Future studies should first evaluate these tissues in laboratory animals and, if the results are positive, design a clinical trial in patients requiring this kind of tissue in order to confirm these results and determine if these tissue substitutes could be clinically useful.…”

“…Fibrin is obtained from human plasma donors, is easily available and can even be of autologous origin, whereas agarose is a natural polymer that has been previously used in food and medical applications, with high biocompatibility and biosafety. In fact, several human tissues and organs have been generated using fibrin‐agarose with encouraging results in vitro and in vivo . For this reason, in the present study we evaluated the usefulness of this biomaterial for generating a substitute of the human bladder mucosa by tissue engineering.…”

“…Tissue biopsies were washed in phosphate‐buffered saline, fragmented in small explants and placed on culture flasks with the epithelial layer in direct contact with the culture surface. Cells grown from these explants were cultured in epithelial culture medium consisting in 75% Dulbecco's modified Eagle's medium and 25% Nutrient Mixture F‐12 Ham with 10% fetal bovine serum, 1% antibiotics/antimycotics, and the following supplements and growth factors: adenine 24 μg/mL, hydrocortisone 0.4 μg/mL, insulin 5 μg/mL, triiodothyronine 1.3 ng/mL, cholera toxin 8 ng/mL and epidermal growth factor 10 ng/mL . To obtain connective stromal cells, the submucosal layer of the tissue biopsies was enzymatically digested with 2 mg/mL of Clostridium histolyticum collagenase I (Gibco BRL Life Technologies, Karlsruhe, Germany) at 37°C for 6 h. Isolated cells were collected by centrifugation and expanded in culture flasks containing basal culture medium (Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics/antimycotics) and using standard cell culture conditions.…”

Microscopic evaluation of transdifferentiation of Wharton’s jelly stem cells to urothelium

“…En este contexto, las células madre mesenquimales de la gelatina de Wharton (HWJSC) podrían representar una fuente alternativa para la construcción de estos tejidos. Las propiedades inmunomoduladoras (4-6), su capacidad de transdiferenciación en los tres linajes embrionarios (7)(8)(9)(10)(11), así como su alta tasa de expansión, convierten a esta estirpe celular en una fuente de células madre idónea para su utilización en protocolos de ingeniería tisular, como se ha demostrado en la construcción de distintos tejidos como son córnea y piel (7)(8)10). El presente trabajo tiene por objeto el estudio de los cambios morfológicos que presentan las HWJSC inducidas hacia urotelio, utilizando un nuevo medio condicionado.…”