Fluorescence-activated cell sorting of PCK-26 antigen-positive cells enables selection of ovine esophageal epithelial cells with improved viability on scaffolds for esophagus tissue engineering

Kofler, Kristina; Ainoedhofer, Herwig; Höllwarth, Michael E.; Saxena, Amulya K.

doi:10.1007/s00383-009-2512-x

Cited by 30 publications

(13 citation statements)

References 22 publications

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“…[41] These proteins can be categorized as either as acidic type-I cytokeratin or as basic/neutral type-II cytokeratin. [42] Both cytokeratin-4 (CK4) and cytokeratin-14 (CK14) are phenotypic differentiation marker proteins of esophageal squamous epithelium. [43] CK14 is characteristically expressed in basal cells while CK4 is typically expressed in 590 P. Kuppan et al…”

Section: Discussionmentioning

confidence: 99%

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-based nanofibrous scaffolds to support functional esophageal epithelial cells towards engineering the esophagus

Kuppan

Sethuraman

Krishnan

2014

Journal of Biomaterials Science, Polymer Edition

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Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and PHBV-gelatin were electrospun to obtain defect-free nanofibers by optimizing various process and solution parameters. Tensile strength, Young's modulus, and wettability of PHBV-gelatin nanofibrous scaffold were determined and compared with PHBV nanofibrous scaffold. Our results demonstrate that PHBV-gelatin nanofibers exhibited higher tensile strength and Young's modulus than the PHBV nanofibers. Human esophageal epithelial cells (HEEpiC) were cultured on PHBV and PHBV-gelatin nanofiber showed better cell proliferation in PHBV nanofibrous scaffold than the PHBV-gelatin scaffold after 7 days of culture. HEEpiC cultured on PHBV and PHBV-gelatin nanofibrous scaffold exhibited characteristic epithelial cobblestone morphology after 3 days of culture. Further, the HEEpiC extracellular matrix (ECM) proteins (collagen type IV and laminin) and phenotypic marker proteins (cytokeratin-4 and 14) expressions were significantly higher in PHBV-gelatin nanofibrous scaffold than the PHBV nanofiber scaffold. However, the long-term stability and functional state of the cells on the PHBV scaffold give it an edge over the blend scaffolds. Thus, PHBV-based nanofibrous scaffolds could be explored further as ECM substitutes for the regeneration of esophageal tissue.

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Section: Discussionmentioning

confidence: 99%

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-based nanofibrous scaffolds to support functional esophageal epithelial cells towards engineering the esophagus

Kuppan

Sethuraman

Krishnan

2014

Journal of Biomaterials Science, Polymer Edition

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“…17 In view of these observations, it seems that communications between different cell types and the matrix are beneficial to tissue remodeling; therefore, the hybrid approach, consisting in assembling acellular scaffolds seeded with epithelial and muscle cells, seems currently the most promising approach. 12,17,39,40 Results of the present study allow us to draw some conclusions regarding the optimal conditions of matrix cell seeding in such a model. A high rate of early myoblast loss was observed when a concentration of 10 6 cells/cm 2 was used.…”

Section: Poghosyan Et Almentioning

confidence: 75%

In Vitro Development and Characterization of a Tissue-Engineered Conduit Resembling Esophageal Wall Using Human and Pig Skeletal Myoblast, Oral Epithelial Cells, and Biologic Scaffolds

Poghosyan

Gaujoux

Vanneaux

et al. 2013

Tissue Engineering Part A

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Introduction: Tissue engineering represents a promising approach for esophageal replacement, considering the complexity and drawbacks of conventional techniques. Aim: To create the components necessary to reconstruct in vitro or in vivo an esophageal wall, we analyzed the feasibility and the optimal conditions of human and pig skeletal myoblast (HSM and PSM) and porcine oral epithelial cell (OEC) culture on biologic scaffolds. Materials and Methods: PSM and HSM were isolated from striated muscle and porcine OECs were extracted from oral mucosa biopsies. Myoblasts were seeded on an acellular scaffold issue from porcine small intestinal submucosa (SIS) and OEC on decellularized human amniotic membrane (HAM). Seeding conditions (cell concentrations [0.5 · 10 6 versus 10 6 cells/cm 2 ] and culture periods [7, 14 and 21 days]), were analyzed using the methyl thiazoltetrazolium assay, quantitative PCR, flow cytometry, and immunohistochemistry. Results: Phenotypic stability was observed after cellular expansion for PSM and HSM (85% and 97% CD56-positive cells, respectively), and OECs (90% AE1/AE3-positive cells). After PSM and HSM seeding, quantities of viable cells were similar whatever the initial cell concentration used and remained stable at all time points. During cell culture on SIS, a decrease of CD56-positive cells was observed (76% and 76% by D7, 56% and 70% by D14, 28% and 60% by D21, for PSM and HSM, respectively). Multilayered surface of a-actin smooth muscle and Desmine-positive cells organized in bundles was seen as soon as D7, with no evidence of cell within the SIS. Myoblasts fusion was observed at D21. Pax3 and Pax7 expression was downregulated and MyoD expression upregulated, at D14.OEC proliferation was observed on HAM with both cell concentrations from D7 to D21. The cell metabolism activity was more important on matrix seeded by 10 6 cells/cm 2 . With 0.5 · 10 6 OEC/cm 2

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“…In order to tissue engineer the esophagus which resembles and functions like the native tissue, a hybrid construct approach that involves assembly of the individual components and amalgamating them is envisaged as the optimal solution [21]. Our efforts in the proliferation of the EEC both in the rodent model as well as the ovine model have demonstrated the identification of the specific cell types required for generation of esophageal epithelium as well as the interaction of EEC on collagen scaffolds to generate the esophageal epithelial component for the hybrid construct [18,[21][22][23]. Our focus now is on the investigation of the smooth muscle cell component for the hybrid construct formation and the reason to perform the present investigation.…”

Section: Discussionmentioning

confidence: 98%

Esophageal smooth muscle cells dedifferentiate with loss of α-smooth muscle actin expression after 8 weeks of explant expansion in vitro culture: Implications on esophagus tissue engineering

et al. 2011

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Background: Esophagus tissue engineering using the hybrid-construct approach involves assembly of the esophageal components to engineer the organ. This study investigated the dedifferentiation and loss in expression of esophageal smooth muscle after explant expansion in culture.Methods: Ovine esophagus smooth muscle cells (OESMC) were sourced from adult biopsies and expanded using the explant culture technique. The explants were maintained under static in vitro tissue culture. Flow cytometry analysis was performed for a-smooth muscle actin (a-SMA) expression at intervals of 4 weeks for up to 8 weeks.Results: After 4 weeks 58.5% OESMC exhibited a-SMA which decreased to 28.5% after 6 weeks in culture. After 8 weeks in culture a mere 1.3% OESMC demonstrated a-SMA expression.Conclusions: OESMC proliferated using the explant technique exhibit a total loss of a-SMA expression after 8 weeks of static in vitro culture. This information is crucial for tissue engineering of the esophagus using the hybridconstruct approach.

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Fluorescence-activated cell sorting of PCK-26 antigen-positive cells enables selection of ovine esophageal epithelial cells with improved viability on scaffolds for esophagus tissue engineering

Cited by 30 publications

References 22 publications

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-based nanofibrous scaffolds to support functional esophageal epithelial cells towards engineering the esophagus

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate)-based nanofibrous scaffolds to support functional esophageal epithelial cells towards engineering the esophagus

In Vitro Development and Characterization of a Tissue-Engineered Conduit Resembling Esophageal Wall Using Human and Pig Skeletal Myoblast, Oral Epithelial Cells, and Biologic Scaffolds

Esophageal smooth muscle cells dedifferentiate with loss of α-smooth muscle actin expression after 8 weeks of explant expansion in vitro culture: Implications on esophagus tissue engineering

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