Obtaining conditional media from Warthon's gelatin mesenchymal/stromal stem cells in a dynamic culture system

Abstract

Introduction. Mesenchymal stem cells (MSCs) have great potential for therapeutic use and exert anti-inflammatory, anti-apoptotic and pro-angiogenic effects through their secretome. MSCs isolated from Wharton's gelatin (WJ-MSCs) have been described as one of the best sources of MSCs and their secretome is rich in bioactive molecules with the ability to maintain cellular and tissue homeostasis for the treatment of pathologies such as inflammation, skin wounds, neurodegenerative disorders, and diabetes.
Objective. To describe a protocol for obtaining WJ-MSCs conditioned media from dynamic cell cultures, considering that the cultivation of MSCs in three-dimensional systems offers advantages over two-dimensional cultures, by better replicating the conditions of the cellular microenvironment in vivo.
Materials and methods: Isolated Wharton gelatin mesenchymal stem cells (WJ-MSCs) were used. The characterization of the WJ-MSCs was carried out by evaluating their immunophenotype by flow cytometry and evaluating multipotency by induction of differentiation to mesenchymal lineages, in vitro.
WJ-MSCs were grown in 2D monolayer cultures and 3D cultures in a 3L Applikon Biotechnology bioreactor using plastic microbeads as microcarriers.
Obtaining the conditioned media was standardized and the detection of 43 angiogenic factors was carried out, using an antibody microarray system for human angiogenesis.
Results. The isolated WJ-MSCs showed typical characteristics of MSCs. Culturing WJ-MSCs in 3D systems facilitated scalability and enhanced secretion of angiogenic factors compared to 2D cultures.
Conclusions. We used 3D and 2D cultures of WJ-MSCs to obtain the secretome of the WJ-MSCs. 3D culture allows obtaining significantly larger volumes of conditioned medium with a potential increased angiogenic capacity.