Paulina Borkowska1, Anna Fila-Danilow2, Monika Paul-Samojedny2, Malgorzata Kowalczyk2, Justyna Hart2, Jakub Ryszawy2, Jan Kowalski2. 1. Department of Medical Genetics, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Sosnowiec, Poland. Electronic address: paubor85@gmail.com. 2. Department of Medical Genetics, School of Pharmacy with the Division of Laboratory Medicine in Sosnowiec, Medical University of Silesia, Sosnowiec, Poland.
Abstract
BACKGROUND: Mesenchymal stem cells (MSCs) are able to differentiate into cells from all three germ layers. The aim of the current work was the differentiation rat MSCs into GABAergic, cholinergic and dopaminergic cells. NEW METHOD: In this paper, we present differentiation cocktails with a hippocampal astrocyte conditioned medium and with a glioblastoma conditioned medium. We wanted to maximize the role of endogenous secreted substances by cells from the central nervous system in both combinations. These modifications create a microenvironment of differentiation that is similar to natural conditions. Moreover, the presence of the Cxcr4 receptor on neuron-like cells was investigated first time. RESULTS: Our results show that a differentiation cocktail with a hippocampal astrocyte conditioned medium is the most effective and that 17% Gad67(+) and 7% Acht(+) cells were observed using this protocol. After differentiation using the glioblastoma conditioned medium, 12% Gad67 (+) was observed. The presence of the Cxcr4 migration receptor on Gad67(+) and Th(+) cells were observed, which might suggest the transplantation potential of differentiated cells. COMPARISON WITH EXISTING METHODS: Our results are slightly lower than those of previous studies but when differences in counting cells is taken into account, a comparison of results is really difficult. CONCLUSIONS: These new differentiation cocktails should be further investigated and in the next experiment only a part of MSCs that expressed the Cxcr4 receptor will be differentiated. We suppose that the Cxcr4(+) cells may differentiate more easily and as a result, we may achieve a homogenous population of one phenotype of neurons.
BACKGROUND: Mesenchymal stem cells (MSCs) are able to differentiate into cells from all three germ layers. The aim of the current work was the differentiation rat MSCs into GABAergic, cholinergic and dopaminergic cells. NEW METHOD: In this paper, we present differentiation cocktails with a hippocampal astrocyte conditioned medium and with a glioblastoma conditioned medium. We wanted to maximize the role of endogenous secreted substances by cells from the central nervous system in both combinations. These modifications create a microenvironment of differentiation that is similar to natural conditions. Moreover, the presence of the Cxcr4 receptor on neuron-like cells was investigated first time. RESULTS: Our results show that a differentiation cocktail with a hippocampal astrocyte conditioned medium is the most effective and that 17% Gad67(+) and 7% Acht(+) cells were observed using this protocol. After differentiation using the glioblastoma conditioned medium, 12% Gad67 (+) was observed. The presence of the Cxcr4 migration receptor on Gad67(+) and Th(+) cells were observed, which might suggest the transplantation potential of differentiated cells. COMPARISON WITH EXISTING METHODS: Our results are slightly lower than those of previous studies but when differences in counting cells is taken into account, a comparison of results is really difficult. CONCLUSIONS: These new differentiation cocktails should be further investigated and in the next experiment only a part of MSCs that expressed the Cxcr4 receptor will be differentiated. We suppose that the Cxcr4(+) cells may differentiate more easily and as a result, we may achieve a homogenous population of one phenotype of neurons.
Authors: Lyudmila M Mezhevikina; Dmitriy A Reshetnikov; Maria G Fomkina; Nurbol O Appazov; Saltanat Zh Ibadullayeva; Evgeniy E Fesenko Journal: Heliyon Date: 2021-03-20