Mesenchymal stem/stromal cells (MSCs) are rare and heterogeneous population of multipotent cells that are present in all human tissues. Perinatal stem cells, or stem cells of birth tissues, include different types of cells, including MSCs, and are derived from perinatal tissues. Most of these cells express certain properties and cellular markers of both multipotency and pluripotency. Due to their genetic stability, they do not form tumors and exhibit immunosuppressive properties through a paracrine communication mechanism. In the case of perinatal stem cells, their easy acquisition, relatively undemanding cultivation in vitro conditions and the possibility of storage in biobanks are crucial. They have the potential to treat various diseases, which is why their development and use in regenerative medicine is rapidly insreasing. The purpose of our research was to compare the isolation efficiency of primary human MSCs derived from four different perinatal tissues of donors after birth. For research purposes, the mothers donated a piece of amniotic membrane (AM), placenta (PL), umbilical cord (POP) and 1 mL of umbilical cord blood (PK). In the study, we examined whether primary cells from these four tissues differ in the time of appearance of adherent cells under in vitro conditions and in their colony-forming ability (CFU-F). The cells were monitored using a phase-contrast version of a light microscope. We recorded the time of appearance of adherent cells and performed the CFU-F test. Based on these results, we evaluated the effectiveness of isolation of primary cells from the investigated tissues. The time to appearance of adherent cells under in vitro conditions differed between primary human MSC obtained from four different perinatal tissues. The PL samples showed a statistically significantly longer time to the appearance of adherent cells compared to samples AM. However, all samples showed the same colony-forming ability and primary cell isolation efficiency. More than half of the samples had ineffective isolation, suggesting the need for improvements in the process of isolating primary cells from these tissues, the culture conditions and the selection of a more suitable cell culture medium. Due to the small number of included donors and the frequent variability of results, a larger number of samples would be needed for a more accurate estimate in order to obtain more reliable and representative data. It would also be useful to analyze the donors, which would allow for a better definition of the target population of pregnant women.
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