Wharton's jelly (WJ) contains mesenchymal stem cells (MSCs) exhibiting broad immunomodulatory properties and differentiation capacity, which makes them a promising tool for cellular therapies. Although the osteogenic, chondrogenic and adipogenic differentiation is a gold standard for proper identification of MSCs, it is important to elucidate the exact molecular mechanisms governing these processes to develop safe and efficient cellular therapies. Umbilical cords were collected from healthy, full-term deliveries, for subsequent MSCs (WJ-MSCs) isolation. WJ-MSCs were cultivated in vitro for osteogenic, chondrogenic, adipogenic and neurogenic differentiation. The RNA samples were isolated and the transcript levels were evaluated using NovaSeq platform, which led to the identification of differentially expressed genes. Expression of H19 and SLPI was enhanced in adipocytes, chondrocytes and osteoblasts, and NPPB was decreased in all analyzed groups compared to the control. KISS1 was down-regulated in adipocytes, chondrocytes, and neural-like cells compared to the control. The most of identified genes were already implicated in differentiation of MSCs; however, some genes (PROK1, OCA2) have not yet been associated with initiating final cell fate. The current results indicate that both osteo- and adipo-induced WJ-MSCs share many similarities regarding the most overexpressed genes, while the neuro-induced WJ-MSCs are quite distinctive from the other three groups. Overall, this study provides an insight into the transcriptomic changes occurring during the differentiation of WJ-MSCs and enables the identification of novel markers involved in this process, which may serve as a reference for further research exploring the role of these genes in physiology of WJ-MSCs and in regenerative medicine.

Cellivia 3 S A 61 623 Poznan Poland

Department of Basic and Preclinical Sciences Institute of Veterinary Medicine Nicolaus Copernicus University in Torun 87 100 Torun Poland

Department of Cancer Immunology Poznan University of Medical Sciences 61 866 Poznan Poland

Department of Histology and Embryology Poznan University of Medical Sciences 60 781 Poznan Poland

Department of Obstetrics and Gynecology University Hospital and Masaryk University 60177 Brno Czech Republic

Department of Toxicology Poznan University of Medical Sciences 60 631 Poznan Poland

Department of Veterinary Surgery Institute of Veterinary Medicine Nicolaus Copernicus University in Torun 87 100 Torun Poland

Division of Anatomy Department of Human Morphology and Embryology Wroclaw Medical University 50 368 Wroclaw Poland

Division of Perinatology and Women's Diseases Poznan University of Medical Sciences 60 535 Poznan Poland

Division of Philosophy of Medicine and Bioethics Poznan University of Medical Sciences 60 806 Poznan Poland

Gene Therapy Laboratory Department of Cancer Diagnostics and Immunology Greater Poland Cancer Centre 61 866 Poznan Poland

Institute of Animal Physiology and Genetics of the Czech Academy of Sciences 27721 Libechov Czech Republic

Physiology Graduate Faculty North Carolina State University Raleigh NC 27695 USA

Prestage Department of Poultry Sciences North Carolina State University Raleigh NC 27695 USA

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