Influence of carrier materials and coatings on retinal pigment epithelium cultivation and functions

Dörschmann, Philipp; Böser, S.; Isik, David; Arndt, Christine; Roider, Johann; Selhuber‐Unkel, Christine; Klettner, Alexa

doi:10.1016/j.exer.2022.109063

Cited by 2 publications

(2 citation statements)

References 56 publications

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“…And our results also confirmed abundant collagen protein in our ADSC-dECM which might contribute to its pro-regenerative function. Additionally, fibronectin and laminin are important ECM protein components involved in cell adhesion and proliferation, which may enhance roughness and facilitate increased cell adhesion and proliferation ( Patten and Wang, 2021 ; Dörschmann et al, 2022 ). And WB analysis also demonstrated increased fibronectin and laminin expressions in our ADSC-dECM.…”

Section: Discussionmentioning

confidence: 99%

Adipose-derived stem cells derived decellularized extracellular matrix enabled skin regeneration and remodeling

Zhang,

Xiang,

Yang

et al. 2024

Front. Bioeng. Biotechnol.

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The tissues or organs derived decellularized extracellular matrix carry immunogenicity and the risk of pathogen transmission, resulting in limited therapeutic effects. The cell derived dECM cultured in vitro can address these potential risks, but its impact on wound remodeling is still unclear. This study aimed to explore the role of decellularized extracellular matrix (dECM) extracted from adipose derived stem cells (ADSCs) in skin regeneration.Methods: ADSCs were extracted from human adipose tissue. Then we cultivated adipose-derived stem cell cells and decellularized ADSC-dECM for freeze-drying. Western blot (WB), enzyme-linked immunosorbent assay (ELISA) and mass spectrometry (MS) were conducted to analyzed the main protein components in ADSC-dECM. The cell counting assay (CCK-8) and scratch assay were used to explore the effects of different concentrations of ADSC-dECM on the proliferation and migration of human keratinocytes cells (HaCaT), human umbilical vein endothelia cells (HUVEC) and human fibroblasts (HFB), respectively. Moreover, we designed a novel ADSC-dECM-CMC patch which used carboxymethylcellulose (CMC) to load with ADSC-dECM; and we further investigated its effect on a mouse full thickness skin wound model.Results: ADSC-dECM was obtained after decellularization of in vitro cultured human ADSCs. Western blot, ELISA and mass spectrometry results showed that ADSC-dECM contained various bioactive molecules, including collagen, elastin, laminin, and various growth factors. CCK-8 and scratch assay showed that ADSC-dECM treatment could significantly promote the proliferation and migration of HaCaT, human umbilical vein endothelia cells, and human fibroblasts, respectively. To evaluate the therapeutic effect on wound healing in vivo, we developed a novel ADSC-dECM-CMC patch and transplanted it into a mouse full-thickness skin wound model. And we found that ADSC-dECM-CMC patch treatment significantly accelerated the wound closure with time. Further histology and immunohistochemistry indicated that ADSC-dECM-CMC patch could promote tissue regeneration, as confirmed via enhanced angiogenesis and high cell proliferative activity.Conclusion: In this study, we developed a novel ADSC-dECM-CMC patch containing multiple bioactive molecules and exhibiting good biocompatibility for skin reconstruction and regeneration. This patch provides a new approach for the use of adipose stem cells in skin tissue engineering.

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Section: Discussionmentioning

confidence: 99%

Adipose-derived stem cells derived decellularized extracellular matrix enabled skin regeneration and remodeling

Zhang,

Xiang,

Yang

et al. 2024

Front. Bioeng. Biotechnol.

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“…VEGF secretion is a natural occurring process at the basal membrane RPE level. We believe that the higher VEGF concentration is a consequence of pronounced maturation of RPE cells over the dDM, since the increase of VEGF stimulates cell attachment and differentiation in order to mediate functional angiogenesis [ 34 ]. Importantly, the phagocytic activity of hESC-RPE cells was enhanced when the cells where cultured over the dDM compared to the control condition, even though not significantly.…”

Section: Discussionmentioning

confidence: 99%

Denuded Descemet’s membrane supports human embryonic stem cell-derived retinal pigment epithelial cell culture

et al. 2023

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Recent clinical studies suggest that retinal pigment epithelial (RPE) cell replacement therapy may preserve vision in retinal degenerative diseases. Scaffold-based methods are being tested in ongoing clinical trials for delivering pluripotent-derived RPE cells to the back of the eye. The aim of this study was to investigate human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells survival and behaviour on a decellularized Descemet’s Membrane (DM), which may be of clinical relevance in retinal transplantation. DMs were isolated from human donor corneas and treated with thermolysin. The DM surface topology and the efficiency of the denudation method were evaluated by atomic force microscope, scanning electron microscopy and histology. hESC-RPE cells were seeded onto the endothelial-side surface of decellularized DM in order to determine the potential of the membrane to support hESC-RPE cell culture, alongside maintaining their viability. Integrity of the hESC-RPE monolayer was assessed by measuring transepithelial resistance. RPE-specific gene expression and growth factors secretion were assessed to confirm maturation and functionality of the cells over the new substrate. Thermolysin treatment did not affect the integrity of the tissue, thus ensuring a reliable method to standardize the preparation of decellularized DM. 24 hours post-seeding, hESC-RPE cell attachment and initial proliferation rate over the denuded DM were higher than hESC-RPE cells cultured on tissue culture inserts. On the new matrix, hESC-RPE cells succeeded in forming an intact monolayer with mature tight junctions. The resulting cell culture showed characteristic RPE cell morphology and proper protein localization. Gene expression analysis and VEGF secretion demonstrate DM provides supportive scaffolding and inductive properties to enhance hESC-RPE cells maturation. Decellularized DM was shown to be capable of sustaining hESC-RPE cells culture, thus confirming to be potentially a suitable candidate for retinal cell therapy.

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Influence of carrier materials and coatings on retinal pigment epithelium cultivation and functions

Cited by 2 publications

References 56 publications

Adipose-derived stem cells derived decellularized extracellular matrix enabled skin regeneration and remodeling

Adipose-derived stem cells derived decellularized extracellular matrix enabled skin regeneration and remodeling

Denuded Descemet’s membrane supports human embryonic stem cell-derived retinal pigment epithelial cell culture

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