Expression Profile of New Marker Genes Involved in Differentiation of Human Wharton's Jelly-Derived Mesenchymal Stem Cells into Chondrocytes, Osteoblasts, Adipocytes and Neural-like Cells.

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.

Apoptosis Related Human Wharton's Jelly-Derived Stem Cells Differentiation into Osteoblasts, Chondrocytes, Adipocytes and Neural-like Cells-Complete Transcriptomic Assays.

Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) exhibit multilineage differentiation potential, adhere to plastic, and express a specific set of surface markers-CD105, CD73, CD90. Although there are relatively well-established differentiation protocols for WJ-MSCs, the exact molecular mechanisms involved in their in vitro long-term culture and differentiation remain to be elucidated. In this study, the cells were isolated from Wharton's jelly of umbilical cords obtained from healthy full-term deliveries, cultivated in vitro, and differentiated towards osteogenic, chondrogenic, adipogenic and neurogenic lineages. RNA samples were isolated after the differentiation regimen and analyzed using an RNA sequencing (RNAseq) assay, which led to the identification of differentially expressed genes belonging to apoptosis-related ontological groups. ZBTB16 and FOXO1 were upregulated in all differentiated groups as compared to controls, while TGFA was downregulated in all groups. In addition, several possible novel marker genes associated with the differentiation of WJ-MSCs were identified (e.g., SEPTIN4, ITPR1, CNR1, BEX2, CD14, EDNRB). The results of this study provide an insight into the molecular mechanisms involved in the long-term culture in vitro and four-lineage differentiation of WJ-MSCs, which is crucial to utilize WJ-MSCs in regenerative medicine.

Ovarian Cancer and Cancer Stem Cells-Cellular and Molecular Characteristics, Signaling Pathways, and Usefulness as a Diagnostic Tool in Medicine and Oncology.

Despite the increasing development of medicine, ovarian cancer is still a high-risk, metastatic disease that is often diagnosed at a late stage. In addition, difficulties in its treatment are associated with high resistance to chemotherapy and frequent relapse. Cancer stem cells (CSCs), recently attracting significant scientific interest, are considered to be responsible for the malignant features of tumors. CSCs, as the driving force behind tumor development, generate new cells by modifying different signaling pathways. Moreover, investigations on different types of tumors have shown that signaling pathways are key to epithelial-mesenchymal transition (EMT) regulation, metastasis, and self-renewal of CSCs. Based on these established issues, new therapies are being investigated based on the use of inhibitors to block CSC growth and proliferation signals. Many reports indicate that CSC markers play a key role in cancer metastasis, with hopes placed in their targeting to block this process and eliminate relapses. Current histological classification of ovarian tumors, their epidemiology, and the most recent knowledge of ovarian CSCs, with particular emphasis on their molecular background, are important aspects for consideration. Furthermore, the importance of signaling pathways involved in tumor growth, development, and metastasis, is also presented.

Human Granulosa Cells-Stemness Properties, Molecular Cross-Talk and Follicular Angiogenesis.

The ovarian follicle is the basic functional unit of the ovary, comprising theca cells and granulosa cells (GCs). Two different types of GCs, mural GCs and cumulus cells (CCs), serve different functions during folliculogenesis. Mural GCs produce oestrogen during the follicular phase and progesterone after ovulation, while CCs surround the oocyte tightly and form the cumulus oophurus and corona radiata inner cell layer. CCs are also engaged in bi-directional metabolite exchange with the oocyte, as they form gap-junctions, which are crucial for both the oocyte's proper maturation and GC proliferation. However, the function of both GCs and CCs is dependent on proper follicular angiogenesis. Aside from participating in complex molecular interplay with the oocyte, the ovarian follicular cells exhibit stem-like properties, characteristic of mesenchymal stem cells (MSCs). Both GCs and CCs remain under the influence of various miRNAs, and some of them may contribute to polycystic ovary syndrome (PCOS) or premature ovarian insufficiency (POI) occurrence. Considering increasing female fertility problems worldwide, it is of interest to develop new strategies enhancing assisted reproductive techniques. Therefore, it is important to carefully consider GCs as ovarian stem cells in terms of the cellular features and molecular pathways involved in their development and interactions as well as outline their possible application in translational medicine.

Human Wharton's Jelly-Cellular Specificity, Stemness Potency, Animal Models, and Current Application in Human Clinical Trials.

Stem cell therapies offer a great promise for regenerative and reconstructive medicine, due to their self-renewal and differentiation capacity. Although embryonic stem cells are pluripotent, their utilization involves embryo destruction and is ethically controversial. Therefore, adult tissues that have emerged as an alternative source of stem cells and perinatal tissues, such as the umbilical cord, appear to be particularly attractive. Wharton's jelly, a gelatinous connective tissue contained in the umbilical cord, is abundant in mesenchymal stem cells (MSCs) that express CD105, CD73, CD90, Oct-4, Sox-2, and Nanog among others, and have the ability to differentiate into osteogenic, adipogenic, chondrogenic, and other lineages. Moreover, Wharton's jelly-derived MSCs (WJ-MSCs) do not express MHC-II and exhibit immunomodulatory properties, which makes them a good alternative for allogeneic and xenogeneic transplantations in cellular therapies. Therefore, umbilical cord, especially Wharton's jelly, is a promising source of mesenchymal stem cells.

[Fetal programming and the etiology of osteoporosis]. / Programowanie plodowe a etiologia osteoporozy.

Osteoporosis is a multifactorial skeletal disorder characterized by low bone mass and microarchitectural deterioration of bone tissue, resulting in increased risk of fracture. Peak bone mass is an important predictor of later risk of osteoporosis. Epidemiological studies revealed that the risk of osteoporosis might be modified by exposure to environmental factors during intrauterine life and early postnatal period. This review summarizes the influence of fetal programming on the development of osteoporosis based on the epidemiological studies and potential mechanisms of epigenetic regulation of gene expression.

The contribution of Hind III C>G PAI-1 gene polymorphism in etiology of recurrent miscarriages.

OBJECTIVES: The goal of the study was to assess the relationship of HindIII C>G PAI-1 gene polymorphism with increased risk of recurrent miscarriages. MATERIAL AND METHODS: A whole of 152 women with a history of at least two miscarriages were classified into analysis. The study group was divided twice (114 subjects with 2 miscarriages and 38 subjects with >3 miscarriages, 123 subjects with miscarriages at <13gw, and 29 subjects with miscarriages in <21 gw). The controls consisted of 180 women with a positive history of at least one pregnancy and birth of a healthy term newborn, and a negative history of miscarriage. The analysed polymorphisms were determined by PCR/RFLP methods. RESULTS: The occurrence of HindIII GG genotype in the whole study group was 25.7% and 20.0% in controls (OR= 1.38, p=0. 14). HindIII G allele was also observed more frequently in the whole study group (45.7% vs. 42.2% in controls, OR=1. 15, p=0.20). The occurrence of HindIII GG genotype was higher in the subgroup of women with >3 miscarriages (31.6% vs. 20.0% in controls, OR= 1.85, p=0.09). HindIII G allele was also noted more frequently in the subgroup of women with >3 miscarriages (50.0% vs. 42.2% in controls, OR=1.37, p=0. 13). A tendency of higher frequency of HindIII GG genotype and HindIII G allele was also noted in the subgroup of patients with miscarriages in the first and second trimester (HindIII GG: 31.0% vs. 20.0% in controls, OR= 1.80, p=O. 14, HindIII G: 51.7% vs. 42.2% in controls, OR=1.4 7, p=0.11). CONCLUSIONS: Mutated HindIII G allele and HindIII GG genotype of HindIII C>G polymorphism probably augment the risk of recurrent miscarriages.

[Polymorphism of bone morphogenetic protein (BMP2) and osteoporosis etiology]. / Polimorfizm bialka morfogenzntycznego kosci (BMP2) a etiologia osteoporozy.

OBJECTIVES: Osteoporosis is a chronic, generalized bone disease conditioned by many factors among which the genetic background plays the significant role. Bone morphogenetic protein (BMP2), a growth factor belong to su- perfamily of TNF- proteins, is actively involved in bone tissue metabolism. BMP2 protein shows the osteoinduction potential and regulates growth of cartilage plate, and the same directly influences the process of osteogenesis. THE AIM: The aim of study was to examine the frequency of genotypes and alleles of 570A>T and 5375G>A of BMP2 gene polymorphisms in population of Polish postmenopausal women, as well as to analyze the relationship between investigated polymorphic variants and bone turnover parameters. MATERIAL AND METHODS: Into the study 117 postmenopausal women, Caucasian race (average age 55,1 years) living in Wielkopolska region were classified. The analysis of 570A>T and 5375G>A BMP2 polymorphisms was performed by polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) while bone mineral density (BMD) was measured by DEXA method. In the research the chosen clinical and bone turnover parameters were analysed. RESULTS: In both 570A>T and 5375G>A BMP2 polymorphisms the similar frequency of genotypes and alleles in investigated groups of postmenopausal women with osteoporosis, osteopenia and in the group with correct T-score were noted. The analysis do not show the relationship of clinical and bone turnover parameters with particular genotypes of BMP2 polymorphisms in women with osteoporosis, osteopenia and in the group with correct T-score. CONCLUSIONS: The research did not confirm directly relationship of 570A>T and 5375G>A BMP2 polymorphisms with osteoporosis development in population of Polish postmenopausal women. The investigation also shows lack of correlation of 570A>T and 5375G>A BMP2 polymorphisms polymorphisms with analysed clinical and bone turnover parameters.

[Genetic polymorphism of the calcitonin receptor gene and bone mineral density in Polish population of postmenopausal women]. / Polimorfizm genu receptora kalcytoniny a gestosc mineralna kosci w populacji kobiet polskich po menopauzie.

INTRODUCTION: In recent years the influence of genetic factors in the pathogenesis of osteopenia and osteoporosis was indicated. The investigations focused on the gene coding for calcitonin receptor. The goal of our analysis was to determine the genotype frequencies of AluI polymorphism of the calcitonin receptor gene (CTR) in the group of Polish postmenopausal women and its possible contribution to osteoporosis development. MATERIAL AND METHODS: 139 postmenopausal women with osteopenia (t-score value from -1.0 to -2.5) (mean age 58.5 +/- 5.9 years, mean age of menopause 49.8 +/- 3.9 years) have been investigated. AluI polymorphism of the CTR gene was determined using PCR/RFLP assay. We have analysed 3 subgroups: CC, CT, and TT. In each subgroup mean weight, height, body mass index (BMI), mean age of menopause and years since menopause (YSM) and parameters of bone turnover: bone mineral density (BMD), t-score, index: young adults (YA) and--age matched (AM) have been analysed. Additionally the group of 138 selected women (mean age 26.5 +/- 4.3 years) as general population has been analysed. RESULTS: In investigated group the frequency of all 3 genotypes was determined as follows: CC: CT : TT = 8.6 : 45.3 : 46.1. Analysing BMD in particular subgroups the higher value for the CT genotype (0.967 +/- 0.161 g/cm2) was found. Similarly t-score (-1.94), YA (80.6%) and AM (90.8%) index were higher in CT genotype carriers. CONCLUSION: Our results suggest possible connection of the AluI polymorphism of the CTR gene with osteopenia and osteoporosis development. To confirm this tendency further investigations in the large number population are necessary.

[PvuII genetic polymorphism of estrogen receptor alpha in the group of postmenopausal women with osteopenia and osteoporosis]. / Znaczenie polimorfizmu Pvuii genu receptora estrogenowego alpha w grupie kobiet po menopauzie z osteopenia oraz osteoporoza.

DESIGN: It was suggested that genetic factors play an important role in the regulation of bone mineral density and in the pathogenesis of bone fracture in osteoporosis. PvuII restriction polymorphism in the intron 1 of the gene coding estrogen receptor alpha (ER-alpha) is indicated to play a significant role in osteopenia and osteoporosis development. The goal of our study was to determine the frequency and the significance of PvuII polymorphism of the ER-alpha gene in the group of postmenopausal women with osteopenia and osteoporosis. MATERIALS AND METHODS: 93 postmenopausal women with osteopenia and osteoporosis (t-score lower than (-1), and 141 healthy postmenopausal women have been investigated for PvuII polymorphism of the ER-a gene using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) assays. RESULTS: We have observed higher frequency of homozygous genotype PP (25.8 vs. 19.8%) and P allele (50.6 vs. 46.1%) in the group of women with low level of t-score. In the osteopenic women (-2.5 < T-score < -1.0) the significant difference in the distribution of t-score index and genotypes has been found. t-score index correlated with body mass index (BMI), mean body weight, BMD aged matched (AM) and BMD young adults (YA) index. The AM and YA index correlated with the number of pregnancies. CONCLUSIONS: The presence of PP genotype and P allele could be connected with higher bone loss and with the development of osteopenia and osteoporosis in postmenopausal women.

[Course of pregnancy, delivery and puerperium in women with Marfan syndrome]. / Przebieg ciazy, porodu i pologu u ciezarnej z zespolem Marfana.

Obstetric course and neonatal outcome in women with Marfan syndrome is the aim of numerous analyses. It is well known that pregnancies complicated by Marfan syndrome increase the risk of maternal morbidity and premature mortality. The main cause of this is cardiovascular abnormalities (aortic root dilatation, aortic dissection, rupture of the aorta, other cardiac abnormalities). We have presented a case history of a pregnant woman with a Marfan syndrome and the review of the literature.