Background. Endometriosis is a chronic and debilitating gynecologic disorder, driven by endocrine and immune dysfunctions, which lead to poor endometrial differentiation and attenuated fertility. Escape from immune surveillance and involvement of inflammatory mechanisms appear to be factors in disease progression. Current diagnostic guidelines for endometriosis still lack an efficient biomarker. Here, we report a study on two previously unexplored factors as potential biomarkers for endometriosis. Methods. A case-control study was performed to evaluate the diagnostic potential of serum CD90 and CD83 levels in endometriosis patients (cases validated by surgical and histological examination) compared to healthy controls. Serum was collected from age-matched females and analyzed by ELISA. Results. Comparison of endometriosis patients to the control group showed significantly elevated levels of serum CD90 ( pg/mL vs. pg/mL; ). A threshold value of 479.4 pg/mL was defined based on the control results, and the diagnostic efficiency of the test was estimated. The obtained sensitivity (70.4%), specificity (92.9%), positive predictive value (90.5%), and negative predictive value (76.5%) rated the test as one with promising diagnostic potential. In contrast, the analysis of serum CD83 levels showed comparable values in both groups, suggesting no association with patient status. Conclusion. Elevated soluble CD90 in human serum is associated with endometriosis, which suggests its putative clinical significance as a biomarker in screening and/or diagnosis of the disease.

Hip replacement is one of the most effective surgical procedures in medicine. Despite the fact that the clinical results of arthroplasty are usually excellent, some implants loosen with time and require revision. To help resolve this problem, research has focused on providing conditions that have a positive biological effect on bone building elements, resulting in a fast, seamless proliferation and differentiation of mesenchymal stem cells (MSC) into osteoblasts. That is why developing an in vitro model for studying MSC functions at the implant–tissue contact zone is essential. The aim of this study was to investigate the proliferation and differentiation capacity of human bone-marrow derived MSC in contact with medical implant-grade Ti-6Al-4V alloy and β-tricalcium phosphate (β-TCP). Human MSC were isolated from bone marrow aspirates obtained from the femoral canal during hip replacement surgeries and seeded on Ti-6Al-4V alloy squared specimens and β-TCP granules. The results indicate that MSC were able to effectively adhere to both biomaterial surfaces and, at the same time, retain their proliferative and osteogenic/adipogenic differentiation potential. We suggest that both Ti-6Al-4V alloy and β-TCP surfaces provide suitable conditions for the proliferation and differentiation of bone marrow MSC and can be used as in vitro models to explore MSC functions at the bone–implant interface.

Granulosa cells represent an essential part of the ovary's somatic compartment that maintains oocyte development and maturation and also provides the hormonal environment necessary to coordinate uterine receptivity with ovulation and, subsequently, to sustain early pregnancy. Primarily due to their major role in the processes of steroidogenesis and folliculogenesis, the study of granulosa cells is likely to be essential for comprehending and elucidating the cellular and molecular mechanisms underlying ovarian development and functions in both physiological and pathological conditions. Density gradient centrifugation is the primary method for the isolation of these cells from follicular fluid. However, this technique is time-consuming and in demand of additional reagents, experience, and dexterity. Furthermore, the limited half-life and proliferative potential of human granulosa cells put additional challenges for their study, making the conventional approaches for cultivation less applicable. Our aim was to develop dependable and effective protocols for the isolation and long-term cultivation of primary granulosa cells while preserving their functional and morphological characteristics. Human granulosa cells were isolated from follicular fluid by centrifugation and treatment with hyaluronidase without using density gradient centrifugation and/or red blood cell lysis buffer procedures. Steroidogenesis, growth dynamics, and the population doubling time of granulosa cells cultivated in a newly designed expansion medium, were determined. The medium consisted of DMEM/F12 supplemented with 5% human serum and 33% human follicular fluid. The isolation technique described in this study ensures the generation of granulosa cells with a very high level of purity. Granulosa cells cultivated in the newly developed expansion medium actively grew without changing morphologically and without losing the ability to produce estradiol and progesterone. Moreover, in this particular expansion medium, granulosa cells demonstrated a significantly increased proliferation rate, with a much shorter population doubling time compared to the control cultures grown in medium containing fetal bovine serum. Here we present a simple, fast, and cost-effective procedure for human granulosa cell isolation, combined with a reliable approach for their rapid growth and long-term cultivation based on a newly formulated expansion medium. Together, these two methods provide a comprehensive system for generating granulosa cell cultures, enabling their extensive study for the purposes of reproductive biology and medicine.

Background: Thin endometrium is a significant cause of infertility due to impaired regeneration, reduced receptivity, and insufficient angiogenesis. Mesenchymal stem cell-derived secretome (MSCsec) is a promising cell-free therapy because it contains bioactive molecules that promote tissue repair. This study evaluated the effects of Wharton’s jelly-derived MSC secretome (WJ-MSCsec) on human endometrial stromal cells (EnSCs) and endothelial cells in vitro. Methods: WJ-MSCs were isolated from umbilical cord tissue, characterized, and cultured under serum-free conditions. The concentrated secretome was analyzed using a human angiogenesis proteome array. EnSCs were isolated from endometrial biopsies. EnSC proliferation and migration in the presence of WJ-MSCsec were assessed using CCK-8 and scratch wound-healing assays. Pro-angiogenic activity of WJ-MSCsec was evaluated using a Matrigel tube formation assay with human umbilical vein endothelial cells (HUVECs). Results: Proteomic analysis of WJ-MSCsec revealed the presence of angiogenic, mitogenic, immunomodulatory, and chemotactic factors. Treatment with WJ-MSCsec significantly enhanced EnSC proliferation and accelerated wound closure. Furthermore, WJ-MSCsec markedly promoted endothelial tube formation, increasing total tube length, junction number, and mesh formation. Conclusions: WJ-MSCsec stimulates EnSC proliferation and migration while enhancing angiogenesis in vitro. WJ-MSCsec represents a promising cell-free therapeutic strategy for endometrial regeneration and reproductive disorders associated with impaired endometrial function.

Background/Objectives: Increasing evidence points to hypoxia and inflammation as two major causes of compromised ovarian function. Increased oxidative stress under hypoxic conditions can damage cellular components, leading to the dysfunction and apoptosis of granulosa cells (GCs). The inflammatory response induced by hypoxia may further impair the function of the ovaries and contribute to the development of premature ovarian insufficiency (POI). In animal models of premature ovarian failure, research has demonstrated that the transplantation of mesenchymal stem cells (MSCs) can enhance reproductive outcomes, increase the number of functioning ovarian follicles, and restore estradiol production. However, the specific mechanisms underlying the observed positive results are not well understood. Methods: The present study provides a comparative analysis of how MSCs influence human GC function under inflammatory and hypoxic conditions, using three different experimental approaches: direct co-culture, indirect co-culture with transwell cell culture inserts, and treatment with MSC-derived conditioned medium (MSCcm). Results: Inflammation significantly suppressed GC estradiol secretion and increased apoptosis. MSCs increased estradiol secretion in normal and hypoxic culture conditions when co-cultured directly with GCs. Our results also showed that, under inflammation, MSCs tended to decrease GC proliferation and that hypoxia alone did not have an effect on GC estradiol secretion or proliferation. Conclusions: The study emphasizes the dual nature of MSCs, which largely determines their effects on other cell types, and the need for the condition-specific optimization of MSC therapies for ovarian regeneration.