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6. Additional file 3 of Efficient differentiation of human embryonic stem cells to retinal pigment epithelium under defined conditions

Author

Limnios, Ioannis J., Chau, Yu-Qian, Skabo, Stuart J., Surrao, Denver C., and O’Neill, Helen C.

Subjects
embryonic structures, sense organs, biological phenomena, cell phenomena, and immunity, equipment and supplies, eye diseases
Abstract

Additional file 3: Fig. S3. Adaptation of the hESC-RPE cell differentiation protocol to xeno-free conditions.

Published
2021
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8. Additional file 1 of Efficient differentiation of human embryonic stem cells to retinal pigment epithelium under defined conditions

Author

Limnios, Ioannis J., Chau, Yu-Qian, Skabo, Stuart J., Surrao, Denver C., and O’Neill, Helen C.

Subjects
embryonic structures, sense organs, biological phenomena, cell phenomena, and immunity, reproductive and urinary physiology
Abstract

Additional file 1: Fig. S1. Initial protocol used for differentiation of hESCs to RPE cells based on embryoid body (EB) formation.

Published
2021
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12. Going beyond RGD : Screening of a cell-adhesion peptide library in 3D cell culture

Author

Sarwat, Mariah, Surrao, Denver C., Huettner, Nick, St John, James A., Dargaville, Tim R., Forget, Aurelien, Sarwat, Mariah, Surrao, Denver C., Huettner, Nick, St John, James A., Dargaville, Tim R., and Forget, Aurelien

Abstract

In tissue engineering, cell-adhesion peptides (CAPs) such as the ubiquitous arginine-glycine-aspartic acid (RGD) sequence have allowed the functionalization of synthetic materials to mimic macromolecules of the extracellular matrix (ECM). However, the variety of ECM macromolecules makes it challenging to reproduce all of the native tissue functions with only a limited variety of CAPs. Screening of libraries of CAPs, analogous to high-throughput drug discovery assays, can help to identify new sequences directing cell organization. However, challenges to this approach include the automation of cell seeding in three dimensions and characterization methods. Here, we report a method for robotically generating a library of 16 CAPs to identify a microenvironment capable of directing a chain-like morphology in olfactory ensheathing cells (OECs), a cell type of particular interest for guiding axon growth in spinal cord injury repair. This approach resulted in the identification of one CAP not previously reported to interact with OECs to direct their morphology into structures suitable for potential axon guidance. The same screening approach should be applicable to any range of cell types to discover new CAPs to direct cell fate or function.

Published
2020

13. Hydrogels with cell adhesion peptide‐decorated channel walls for cell guidance

Author

Wang, Shuang, Sarwat, Mariah, Wang, Peng, Surrao, Denver C., Harkin, Damien G., St John, James A., Bolle, Eleonore C.L., Forget, Aurelien, Dalton, Paul D., Dargaville, Tim R., Wang, Shuang, Sarwat, Mariah, Wang, Peng, Surrao, Denver C., Harkin, Damien G., St John, James A., Bolle, Eleonore C.L., Forget, Aurelien, Dalton, Paul D., and Dargaville, Tim R.

Abstract

A method is reported for making hollow channels within hydrogels decorated with cell-adhesion peptides exclusively at the channel surface. Sacrificial fibers of different diameters are used to introduce channels within poly(ethylene glycol) hydrogels crosslinked with maleimide-thiol chemistry, which are backfilled with a cysteine-containing peptide solution which is conjugated to the lumen with good spatial efficiency. This allows for peptide patterning in only the areas of the hydrogel where they are needed when used as cell-guides, reducing the amount of required peptide 20-fold when compared to bulk functionalization. The power of this approach is highlighted by successfully using these patterned hydrogels without active perfusion to guide fibroblasts and olfactory ensheathing cells—the latter having unique potential in neural repair therapies.

Published
2020

21. Can Microcarrier-Expanded Chondrocytes Synthesize Cartilaginous Tissue In Vitro?

Author

Surrao, Denver C., Khan, Aasma A., McGregor, Aaron J., Amsden, Brian G., and Waldman, Stephen D.

Abstract

Tissue engineering is a promising approach for articular cartilage repair; however, it is challenging to produce adequate amounts of tissue in vitrofrom the limited number of cells that can be extracted from an individual. Relatively few cell expansion methods exist without the problems of de-differentiation and/or loss of potency. Recently, however, several studies have noted the benefits of three-dimensional (3D) over monolayer expansion, but the ability of 3D expanded chondrocytes to synthesize cartilaginous tissue constructs has not been demonstrated. Thus, the purpose of this study was to compare the properties of engineered cartilage constructs from expanded cells (monolayer and 3D microcarriers) to those developed from primary chondrocytes. Isolated bovine chondrocytes were grown for 3 weeks in either monolayer (T-Flasks) or 3D microcarrier (Cytodex 3) expansion culture. Expanded and isolated primary cells were then seeded in high density culture on Millicell™ filters for 4 weeks to evaluate the ability to synthesize cartilaginous tissue. While microcarrier expansion was twice as effective as monolayer expansion (microcarrier: 110-fold increase, monolayer: 52-fold increase), the expanded cells (monolayer and 3D microcarrier) were not effectively able to synthesize cartilaginous tissue in vitro. Tissues developed from primary cells were substantially thicker and accumulated significantly more extracellular matrix (proteoglycan content: 156%–292% increase; collagen content: 70%–191% increase). These results were attributed to phenotypic changes experienced during the expansion phase. Monolayer expanded chondrocytes lost their native morphology within 1 week, whereas microcarrier-expanded cells were spreading by 3 weeks of expansion. While the use of 3D microcarriers can lead to large cellular yields, preservation of chondrogenic phenotype during expansion is required in order to synthesize cartilaginous tissue.

Published
2011
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22. Design and characterization of a biodegradable composite scaffold for ligament tissue engineering.

Author

Hayami JW, Surrao DC, Waldman SD, and Amsden BG

Subjects
Animals, Biomarkers metabolism, Cattle, Cell Culture Techniques methods, Cells, Cultured, Chitosan chemistry, Chitosan metabolism, Cross-Linking Reagents chemistry, Cross-Linking Reagents metabolism, Dioxanes chemistry, Dioxanes metabolism, Extracellular Matrix chemistry, Fibroblasts cytology, Fibroblasts metabolism, Guided Tissue Regeneration instrumentation, Guided Tissue Regeneration methods, Humans, Ligaments cytology, Ligaments metabolism, Materials Testing, Methacrylates chemistry, Methacrylates metabolism, Polyesters chemistry, Polyesters metabolism, Absorbable Implants, Biocompatible Materials chemistry, Biocompatible Materials metabolism, Hydrogels chemistry, Hydrogels metabolism, Ligaments chemistry, Tissue Engineering instrumentation, Tissue Engineering methods, Tissue Scaffolds chemistry
Abstract

Herein we report on the development and characterization of a biodegradable composite scaffold for ligament tissue engineering based on the fundamental morphological features of the native ligament. An aligned fibrous component was used to mimic the fibrous collagen network and a hydrogel component to mimic the proteoglycan-water matrix of the ligament. The composite scaffold was constructed from cell-adherent, base-etched, electrospun poly(epsilon-caprolactone-co-D,L-lactide) (PCLDLLA) fibers embedded in a noncell-adherent photocrosslinked N-methacrylated glycol chitosan (MGC) hydrogel seeded with primary ligament fibroblasts. Base etching improved cellular adhesion to the PCLDLLA material. Cells within the MGC hydrogel remained viable (72 +/- 4%) during the 4-week culture period. Immunohistochemistry staining revealed ligament ECM markers collagen type I, collagen type III, and decorin organizing and accumulating along the PCLDLLA fibers within the composite scaffolds. On the basis of these results, it was determined that the composite scaffold design was a viable alternative to the current approaches used for ligament tissue engineering and merits further study., ((c) 2009 Wiley Periodicals, Inc.)

Published
2010
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