Cynthia G Trejo-Iriarte, Miguel A Ortega, Ángel Asúnsolo, José F Gómez-Clavel, Alejandro García Muñoz, Melchor Álvarez- Mon, Julia Buján, Julio Acero, Natalio García-Honduvilla
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引用次数: 4
Abstract
Mesenchymal cells (MSCs) are considered to be cellular populations of common embryological origin. For clinical research applications, MSCs are expanded and increased with cells obtained from a primary culture. By extracting cells from tissue and encouraging them to reproduce, the stem cell population ends up dominating the culture due to a high proliferation rate and self-renewal. The first subcultures between the third and sixth are chosen in order to obtain the maximum number of cells with optimal differentiation capacity. However, few studies have reported long-term cultivation of MSCs. The objective of this study was to advance the knowledge on the characteristics of MSCs by assessing their capacity for self-renewal and phenotypic maintenance beyond 50 cell subcultures, which is defined as the normal limit for cellular survival. Rat subcutaneous adipose tissue was the source of mesenchymal adipose stem cells (MASCs) cultured over 175 subcultures. Early 1 to 5 and late 25 to 30 subcultures were used to induce cellular differentiation to become adipogenic, chondrogenic and osteogenic connective tissue cells. MASCs characteristics were studied using flow cytometry, transmission electron microscopy (TEM), and immunohistochemical and reverse transcription polymerase chain reaction (RT-qPCR) assays. The MASCs maintained cell differentiation capacity for more than 30 subcultures but lost potentiality starting at 60 up to 175 subcultures. MASCs showed the embryonic phenotypes OCT3/4 and Nanog indefinitely, and developed compensatory mechanisms, such as autophagy, to achieve cell survival over a long time period. Therefore, long-term subcultures showed that MASCs could maintain their potential for clinical research use.
期刊介绍:
The official journal of the National Society for Histotechnology, Journal of Histotechnology, aims to advance the understanding of complex biological systems and improve patient care by applying histotechniques to diagnose, prevent and treat diseases.
Journal of Histotechnology is concerned with educating practitioners and researchers from diverse disciplines about the methods used to prepare tissues and cell types, from all species, for microscopic examination. This is especially relevant to Histotechnicians.
Journal of Histotechnology welcomes research addressing new, improved, or traditional techniques for tissue and cell preparation. This includes review articles, original articles, technical notes, case studies, advances in technology, and letters to editors.
Topics may include, but are not limited to, discussion of clinical, veterinary, and research histopathology.