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Special Issue "Tissue Engineering and Cell Therapy: Recent Progress"

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1. Department of Surgery, Faculty of Medicine, Université Laval, Québec, QC G1V 0A6, Canada
2. Regenerative Medicine Division, CHU de Québec—Université Laval Research Centre, Québec, QC G1J 1Z4, Canada
3. Université Laval Research Center on Experimental Organogenesis/LOEX, Québec, QC G1J 1Z4, Canada
Interests: epithelial stem cells; tissue engineering; regenerative medicine; cell therapy; skin
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Special Issue Information

Dear Colleagues,

The clinical need to replace wounded, damaged or pathological tissues and organs has fostered the development of tissue engineering and cellular therapies. Innovative research on biomaterials, extracellular matrix and cells has resulted in tissue substitutes or cell products that are designed to improve, repair or replace several tissues and organs. This Special Issue will gather studies on the recent advances in this area. It will feature research that reports such progress, including fundamental studies on reconstructed tissues in vitro or translational studies and pre-clinical testing in vivo.

Prof. Dr. Lucie Germain
Guest Editor

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Keywords

biomaterials and cell sources for tissue engineering and cell therapy
tissue engineering of substitutes in vitro
functional testing of tissues reconstructed in vitro
extracellular matrix
mechanical characteristics
biological and histological characterization
translational research

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Research

Open AccessArticle

Comparison of Two Human Skin Cell Isolation Protocols and Their Influence on Keratinocyte and Fibroblast Culture

Álvaro Sierra-Sánchez

Martin A. Barbier

Brice Magne

Danielle Larouche

Salvador Arias-Santiago

and

Lucie Germain

Int. J. Mol. Sci. 2023, 24(19), 14712; https://doi.org/10.3390/ijms241914712 - 28 Sep 2023

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Abstract

For the development of advanced therapies, the use of primary cells instead of cell lines is preferred. The manufacture of human tissue-engineered skin substitutes requires efficient isolation and culture protocols allowing a massive expansion of the cells in culture from an initial specimen of a minimal size. This study compared two skin cell isolation protocols, routinely applied in two clinical laboratories. Epithelial (keratinocytes) and dermal (fibroblasts) cells were isolated and cultured from three human skin biopsies (N = 3). The two-step digestion protocol (LOEX-Protocol) firstly used thermolysin to enzymatically disrupt the dermal–epidermal junction while, for the one-step digestion protocol (UPCIT-Protocol), mechanical detachment with scissors was applied. Then, the epidermal and dermal layers were digested, respectively, to achieve cell isolation. The cell size, viability, yield and growth were analyzed over five passages (P). The colony-forming efficiency (CFE) and Keratin 19 (K19) expression of epithelial cells were also assessed after P0 and P1. Regarding the dermal cells, no significant differences were observed in the tested parameters of isolation and culture. However, for the epithelial cells, viability was higher (93% vs. 85%) and the number of cells extracted per cm² of skin was 3.4 times higher using the LOEX-Protocol compared to the UPCIT-Protocol. No significant difference was observed for any parameter once the keratinocytes were cultured from P1 to P4. The CFE and K19 expression decreased from P0 to P1 in both protocols, probably due to the culture process. This study shows that both protocols enable the efficient isolation of skin dermal and epithelial cells and subsequent culture to produce grafts destined for the treatment of patients. Full article

(This article belongs to the Special Issue Tissue Engineering and Cell Therapy: Recent Progress)

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Open AccessArticle

Production of Tissue-Engineered Skin Substitutes for Clinical Applications: Elimination of Serum

and

Int. J. Mol. Sci. 2023, 24(16), 12537; https://doi.org/10.3390/ijms241612537 - 08 Aug 2023

Viewed by 793

Abstract

Tissue-engineered skin substitutes (TESs) are used as a treatment for severe burn injuries. Their production requires culturing both keratinocytes and fibroblasts. The methods to grow these cells have evolved over the years, but bovine serum is still commonly used in the culture medium. Because of the drawbacks associated with the use of serum, it would be advantageous to use serum-free media for the production of TESs. In a previous study, we developed a serum-free medium (Surge SFM) for the culture of keratinocytes. Herein, we tested the use of this medium, together with a commercially available serum-free medium for fibroblasts (Prime XV), to produce serum-free TESs. Our results show that serum-free TESs are macroscopically and histologically similar to skin substitutes produced with conventional serum-containing media. TESs produced with either culture media expressed keratin 14, Ki-67, transglutaminase 1, filaggrin, type I and IV collagen, and fibronectin comparably. Mechanical properties, such as contraction and tensile strength, were comparable between TESs cultured with and without serum. Serum-free TESs were also successfully grafted onto athymic mice for a six-month period. In conclusion, Surge SFM and Prime XV serum-free media could be used to produce high quality clinical-grade skin substitutes. Full article

(This article belongs to the Special Issue Tissue Engineering and Cell Therapy: Recent Progress)

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Special Issue "Tissue Engineering and Cell Therapy: Recent Progress"

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