Volume 8 (2020): Issue 3 (December 2020)

The placenta is a part of feto-maternal unit that develops from the maternal decidua basalis and fetal-derived trophoblast cells. The regulation of its early development is extremely intricate, albeit the elusive trophoblast stem cells (TSCs) are thought to give rise to the fetal part of the placenta. TSCs may be isolated in both animal and human models. In detail, TSCs can be efficiently obtained from the early conceptus tissues – blastocysts or early placental tissue. The isolation of murine TSCs pave the way for analyses of human trophoblast cell lineages. Both human and animal stem cells retain similar characteristic properties – the ability for unrestricted self-renewal and differentiation into all trophoblast cell lines. Nevertheless, there are some essential differences across the various species which are especially pronounced when pertaining to their distinct optimal cell culture requirements. Moreover, there are several crucial discrepancies in the stemness marker gene transcription profiles between human and murine TSCs models. In vitro TSC models can be adapted to the elucidation of the pathophysiology of various reproductive complications. For instance, their properties may illustrate the conditions observed during the implantation or simulate the state of abnormal placentation. Observations gained from the experimental studies could potentially explain the cause of some cases of infertility, preeclampsia, and fetal growth abnormalities.

Running title: Update on the trophoblast stem cells

Exosomes belong to structures called extracellular vesicles (EVs). These spherical units, secreted by most eukaryotic cells, attracted significant interest among researchers in recent years. Exosomes undergo secretion from almost all types of mammalian cells, including dendritic cells, B cells, epithelial cells, mastocytes, reticulocytes, platelets, T cells, mesenchymal stem cells, adipocytes, bone marrow-derived stem cells, embryonic stem cells, fibroblasts, cardiac myocytes, endothelial cells, oligodendrocytes, astrocytes, microglia, neurons, neural stem cells, hepatocytes, lung spheroid cells, as well as tumor cells. Exosomes have several features that enable many methods of their isolation from biological material. Furthermore, physicochemical properties such as size, mass, density, or the ability to interact with specific proteins allowed for the development and advance of several effective methods. Work on exosomes’ recovery and purity made it possible to most effectively determine their isolation methods’ efficiency and accuracy. A common ground for the researchers’ interest in exosomal analyses is the role of exosomes as carriers of disease biomarkers. It has been suggested that exosomes can be used in vaccine development and other immunological-related purposes, as one of their characteristics is the ability to present antigens. Moreover, exosomes have a long half-life. As the human body does not perceive them as foreign bodies, they can penetrate cell membranes and target specific cells, making them even better candidates for the applications mentioned above. Therefore, the following review deals with the nature of exosomes, as well as various methods of their isolation and use in medicine.

Running title: Current application of exosomes in medicine

In the female reproductive tract, reactive oxygen species (ROS) may exert physiological and pathophysio-logical effects. Although ROS play an essential role as the signaling molecules, their excessive accumulation contributes to the pathogenesis of many reproductive processes. In the ovarian follicle, ROS affect multiple physiological processes, including oocyte maturation and fertilization. However, a lack of studies showing to which extend ovarian granulosa and cumulus cells can contribute to the development of oxidative stress within the ovarian follicle. In the presented research, the extracellular ROS accumulation level was investigated using GCs and CCs primary in vitro cultures. The obtained results demonstrated a steady decrease in extracellular ROS level during GCs primary culture. By contrast, ROS concentration in CCs conditioned medium increased gradually between the first and the seventh days of culture. The observed changes may reflect the proliferation status and metabolic activity of GCs and CCs during in vitro culture. Additionally, the elevated ROS level at respective points of time could occur as a consequence of culture in atmospheric oxygen. The distinct function and localization within the ovarian follicle may explain the differences between GCs and CCs oxygen metabolism.

Running title: Reactive oxygen species in primary culture of human follicular cells

Several genes, namely CD44, CD90, CD105 and PCNA may be important in differentiation of porcine mucosa cell cultures. These genes are, inter alia, responsible for cell adhesion to extracellular matrix and its constituent secretion, cytoskeleton organization, epithelial to mesenchymal transition or proper course of DNA replication. A total of 20 pubertal crossbred Landrace gilts bred on commercial farms were used to produce buccal mucosa cultures, which were harvested on the 7th, 15th and 30th day after initiation of the culture. Expression levels of CD44, CD90, CD105 and PCNA were evaluated employing Real-Time Quantitative Polymerase Chain Reaction. CD44, CD90 and PCNA showed an unchanged expression pattern. Expression of CD44 on day 7 was the highest of all factors measured. The greatest difference between the measurement on 7th and 30th day was found in the PCNA gene. These results broaden the understanding of the transcriptome changes in porcine buccal mucosa cells for the duration of in vitro cultivation. Nevertheless, it is very important to consider that the in vitro conditions do not fully reflect the changes taking place in the living organism. It appears that tissues of the oral cavity possess high regenerative potential, and constitute suitable model for wound healing investigation.

Running title: Confirmation of differentiation clusters’ and endoglin markers preset in porcine buccal mucosa cells

Stem cells and their usage for a long time are thought to be the future and hope in modern medicine. In this review we summarize development in science and bioengineering in this field. Opening with a description of newly discovered and studied sources of stem cells acquisition we present scientific methods progress and their application like 3D printing or transdifferentiation mode of action and results of these techniques. Technologies of genome editing like transcription activator-like effector nuclease, zinc-finger nucleases, or CRISPR Cas9 are also presented. In disease treatment and tissue reconstruction stem cells have proved to be effective most times due to great proliferation and differentiation potentials in presented in this summary pre-clinical and clinical studies for diseases like peripheral nerve palsy, myocardial infarction and heart ischemic disease and corneal wound healing.

Running title: Current stem cells technologies used in medicine