Gelatin-Methacryloyl (GelMA) Formulated with Human Platelet Lysate Supports Mesenchymal Stem Cell Proliferation and Differentiation and Enhances the Hydrogel's Mechanical Properties.

Wiebke Handke,Iliyana Pepelanova,Thomas Scheper,Axel Seltsam,Luise Birnstein,Jessica Rach,Marline Kirsch,Antonina Lavrentieva

doi:10.3390/bioengineering6030076

Wiebke Handke, Iliyana Pepelanova + Show 6 more

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https://doi.org/10.3390/bioengineering6030076

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Abstract

Three-dimensional (3D) cell culture is a major focus of current research, since cultivation under physiological conditions provides more reliable information about in vivo cell behavior. 3D cell cultures are used in basic research to better understand intercellular and cell-matrix interactions. Moreover, 3D cell culture plays an increasingly important role in the in vitro testing of bioactive substances and tissue engineering. Gelatin-methacryloyl (GelMA) hydrogels of different degrees of functionalization (DoFs) are a versatile tool for 3D cell culture and related applications such as bioprinting. Human platelet lysate (hPL) has already demonstrated positive effects on 2D cell cultures of different cell types and has proven a valuable alternative to fetal calf serum (FCS). Traditionally, all hydrogels are formulated using buffers. In this study, we supplemented GelMA hydrogels of different DoF with hPL during adipose tissue-derived mesenchymal stem cell (AD-MSCs) encapsulation. We studied the effect of hPL supplementation on the spreading, proliferation, and osteogenic differentiation of AD-MSCs. In addition, the influence of hPL on hydrogel properties was also investigated. We demonstrate that the addition of hPL enhanced AD-MSC spreading, proliferation, and osteogenic differentiation in a concentration-dependent manner. Moreover, the addition of hPL also increased GelMA viscosity and stiffness.

Highlights

Three-dimensional (3D) cell culture systems provide a more physiological environment for cells in comparison to traditional 2D cell cultures in terms of cell-cell and cell-matrix interactions and diffusion behavior
To evaluate the influence of human platelet lysate (hPL) on 3D-cultivated adipose tissue-derived mesenchymal stem cell (AD-Mesenchymal stem cells (MSCs)), cells were encapsulated in three different GelMA-hydrogels of 50%, 70%, and 95% degrees of functionalization (DoF)
Each GelMA solution was prepared in PBS with the addition of 0%, 2.5%, or 50% hPL (% v/v) or by adding hPL only (100% hPL), prior to the addition of the photoinitiator and the polymerization of the hydrogels

Summary

Introduction

Three-dimensional (3D) cell culture systems provide a more physiological environment for cells in comparison to traditional 2D cell cultures in terms of cell-cell and cell-matrix interactions and diffusion behavior. No physiological gradients of signaling molecules, metabolites, and oxygen can be created in 2D culture systems For these reasons, 3D cultures are increasingly used in basic research, drug screening, toxicity studies, and tissue engineering (TE) [1,2,3,4]. Gelatin-methacryloyl (GelMA) provides a broad spectrum of tunable parameters, which lead to the creation of hydrogel networks with different stiffness and pore architectures [10,11,12,13]. These hydrogels can serve as a versatile platform for 3D cell culture research and TE. Engineered polymers like hydrogels represent a valuable alternative to decellularized tissues

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