Inexpensive production of near-native engineered stromas

Abstract

Although the self-assembly approach is an efficient method for the production of engineered physiological and pathological tissues, avoiding the use of exogenous materials, it nevertheless remains expensive and requires dexterity, which are features incompatible with large-scale production. We propose a modification to this technique to make easier the production of mesenchymal compartment, to reduce the cost and to improve the histological quality of the self-assembled tissues. The stroma produced by this novel approach allowed epithelial cell differentiation, resulting in a pseudostratified epithelium that shared several features with native tissues. The incorporation of endothelial cells in the reconstructed mesenchyme formed a three-dimensional capillary-like network, positive for CD31 and von Willebrand factor and surrounded by NG2 positive cells. It could limit self-contraction of the resulting tissue by recruiting α-Smooth Muscle Actin positive cells. With this new technique, which is relatively inexpensive and easy to use in a research laboratory set-up, near-native stromas can now be produced with minimal handling time. Copyright © 2015 John Wiley & Sons, Ltd.

Tissue-engineered 3D melanoma model with blood and lymphatic capillaries for drug development

“…This low throughput model, by recapitulating key features of epidermal differentiation and capillary assembly, can be an excellent tool to complete and confirm data obtained from high throughput screening using spheroids alone 46,60 . The production time of the dermal compartment can be further reduced by using reseeding methods 61 . Being produced in standard 12 well- and 6 well culture plates, tissues can easily be adapted for partially automatized production, allowing a scale-up and a more high throughput-like use.…”

Urothelial cell expansion and differentiation are improved by exposure to hypoxia

“…Urothelial cells were extracted from a human urological tissue biopsy and were cultured as previously described (Chabaud et al , ). UCs were seeded at a density of 5 × 10 5 cells in 75‐cm 2 culture flasks with 1.5 × 10 5 irradiated murine S3T3 as a feeder layer in DMEM with Ham's F12 in a 3:1 proportion (Flow Laboratory, Mississauga, Ontario, Canada) containing 10% fetal bovine serum (Hyclone), 5 mg/ml insulin (Sigma), 0.4 mg/ml hydrocortisone (Calbiochem, San Diego, CA, USA), 10 −10 m cholera toxin (Sigma), 10 μg/ml epidermal growth factor (Austral Biologicals, San Ramon, CA, USA), 100 U/ml penicillin, 25 mg/ml gentamicin and buffered with sodium bicarbonate (UC media).…”

Natural marine sponges for bone tissue engineering: The state of art and future perspectives

“…In this context, tissue engineering (TE) has been emerging as a promising field to develop appropriate models and technologies to promote regeneration of human tissues or to replace damaged or defective organs . TE strategies include cell‐based therapies to create new tissues via cellular biochemical machinery stimulation . Furthermore, the implantation of nonliving components, such as three‐dimensional (3D) scaffolds or matrices, has been widely studied, especially to support cell attachment and growth .…”

“…6 TE strategies include cell-based therapies to create new tissues via cellular biochemical machinery stimulation. 7,8 Furthermore, the implantation of nonliving components, such as threedimensional (3D) scaffolds or matrices, has been widely studied, especially to support cell attachment and growth. 9 Indeed, 3D scaffolds are one of the most promising experimental approaches for regenerating the native structural Correspondence to: R. N. Granito (e-mail: re_neves@yahoo.com.br) and functional properties of living tissues, providing a 3D physical support for in vitro cell culture as well as a matrix for tissue regeneration in vivo.…”

“…9 Indeed, 3D scaffolds are one of the most promising experimental approaches for regenerating the native structural Correspondence to: R. N. Granito (e-mail: re_neves@yahoo.com.br) and functional properties of living tissues, providing a 3D physical support for in vitro cell culture as well as a matrix for tissue regeneration in vivo. 7,8 For a better biological performance, scaffolds should present specific characteristics, such as appropriate pore size and porosity, which are essential for cells to migrate and to grow. 10 Another characteristic of an adequate scaffold is a proper biodegradability, which allows the scaffold to be replaced by the new tissue.…”