Aim: Adding platelet-rich plasm (PRP) or mesenchymal stem cells are the most accepted methods to increase fat graft’s permanence. However, there is no consensus on timing and whether the effect of stem cells or PRP is observed more in the recipient area or in the donor area. It is aimed to present the application method and localization of PRP to be combined with fat graft to increase the survival. Material and Methods: in vitro part: Fat grafts were kept in a medium containing PRP. Cell output from the fat graft to the Petri dishes was examined every day. The time to recruit the maximum number of stem cells to the medium was determined (day 10). in vivo part: Eight group of rats were received PRP in different concentrations either to recipient or donor areas of fat grafts. After ten days, fat grafts were transferred to the recipient area. Ninety days after the transfer, histopathological examination and immunohistochemical staining were performed. Results: The group which had received the full-dose PRP to the recipient area showed the highest cellular integrity and vascularity (p< 0.05). Vascularity was superior in the half-dose PRP group compared to the control group (p< 0.05); cellular integrity did not increase. Donor site groups did not show increase in cellular integrity. Conclusion: Although PRP starts to increase fat graft permanence by increasing vascularity, it acts by preserving the cellular integrity of the fat cells as the concentrations increase. The PRP injection to the recipient area 10 days before fat grafting may provide higher survival rates.

Objective: Mesenchymal stem cells are used in various fields, such as cellular therapy, regeneration, or tissue engineering. It has been shown that they exhibit many protective factors and also work as a modulating chief within the region in which they are administered. There are studies on both the therapeutic and neuroprotective effects of brain-derived neurotrophic factor. Also, there are many studies on the improvement of culture conditions for in vitro reproduction of mesenchymal stem cells, which can be obtained from many sources in various bodies, such as adipose tissue and Wharton’s jelly. Improving and standardizing these culture conditions will increase the effectiveness and reliability of stem cell therapies. Studies evaluating many culture conditions, such as O2 level, type of medium, monolayer culture, and the transition from in vitro 3D models, are ongoing. Materials and Methods: In our study, groups were formed by using stem cells originating from adipose tissue and Wharton’s jelly. Stem cell cultures were made using Hillex-II and Pronectin-F microcarriers. Cell culture O2 level was adjusted as 1% and 5% for each group separately. Enzyme-linked immunosorbent assay was used to analyze brain-derived neurotrophic factor levels in stem cell culture fluid. Results: The highest brain-derived neurotrophic factor level in mesenchymal stem cells culture medium was observed in an adipose-derived stem cell culture with an in vitro fertilization (non-treated) dish, using a Hillex microcarrier in a 1% O2 microenvironment. Conclusion: As a result of our observations, we think that cells could exhibit greater therapeutic potential in a dynamic adhesion environment. Cite this article as: Çiçek G, Çiçek M, Utlu Özen E, Aktan TM, Özdengül F, Duman S. Brain-derived neurotrophic factor levels in mesenchymal stem cell culture fluid under different culture conditions. Eurasian J Med 2023;55(1):25-31.

Hypoxic preconditioning (HP) is a promising approach to enhance the therapeutic efficacy of mesenchymal stem cells (MSCs) by modulating their oxidative stress response, metabolic activity, and secretome composition. Conditioned media (CM) obtained from MSCs cultured under hypoxia contains bioactive molecules and extracellular vesicles (EVs) that support regenerative processes. However, the effects of varying oxygen levels on the redox status and physicochemical characteristics of MSC-derived CM remain incompletely understood. This study aimed to investigate how two physiologically relevant oxygen concentrations (1% and 5%) influence oxidative stress parameters and nanoparticle features in Wharton’s jelly-derived MSC (WJ-MSC)-conditioned media. Cells were cultured under 1% or 5% O₂ and subjected to serum starvation for 48 or 72 h. CM samples were analyzed for total oxidant status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI). Nanoparticle size and zeta potential were evaluated using dynamic light scattering (DLS), and HIF-1α levels were quantified via ELISA. Results showed that CM from 1% O₂ cultures exhibited significantly higher oxidative stress, with elevated TOS and OSI values and reduced TAS levels, particularly after 72 h. Nanoparticle size was initially larger under 1% O₂ but decreased with time, whereas 5% O₂ supported more stable size profiles. Zeta potential measurements revealed more negative values under 5% O₂, indicating greater colloidal stability. HIF-1α expression markedly increased under 1% O₂, confirming hypoxia-induced cellular adaptation. In conclusion, this study demonstrates that graded hypoxia distinctly modulates oxidative stress and nanoparticle characteristics in MSC-derived CM. These findings provide a basis for optimizing hypoxic preconditioning protocols to improve the quality and therapeutic potential of acellular MSC-based therapies.

Amaç: Bu çalışmanın amacı, hastanemize infertilite nedeniyle başvuran erkeklere yardımcı üreme tekniklerinden önce uygun genetik danışmanlık verebilmek için, azospermi ve/veya oligozoospermi etiyolojisine yönelik standart sitogenetik yöntemler ve Y kromozom mikrodelesyon analizleri ile hem majör kromozom anomalilerinin hem de Y kromozomu mikrodelesyonlarının sıklığı ve tiplerini araştırmaktır. Gereç ve Yöntemler: Çalışmamıza 2017-2020 yılları arasında erkek infertilitesi nedeniyle Kanuni Sultan Süleyman Eğitim ve Araştırma hastanemize başvuran toplam 437 hasta dâhil edildi. Tüm hastalar spermiogram, hormonal profil, kromozom analizi ve Y mikrodelesyon analizleri doğrultusunda değerlendirildi. Bulgular: Çalışmamızda toplam 437 hastanın 42’sinde (%9,6) kromozomal anomaliler tespit edildi. En sık görülen kromozomal anomali 47,XXY(Klinefelter sendromu) idi. 5 hastamızda dengeli translokayonlar vardı. 1 hastada ise marker kromozom tespit edildi. Geriye kalan 395 hastanın kromozom analizi normaldi. 44 hastamızda (%10,06) ise AZF genleri üzerinde çeşitli y-kromozomu mikrodelesyonları saptandı. 1 hastada AZFa delesyonu, 4 hastada AZFb+c delesyonu, 17 hastada AZF-c gr/gr delesyonu, 2 hastada komplet AZFa+b+c delesyonu, 2 hastada komplet AZFc delesyonu, 1 hastada parsiyel AZFb delesyonu, 9 hastada kısmi AZFb+c delesyonu, 8 hastada parsiyel AZFc delesyonu tespit edildi. Geriye kalan 393 hastada herhangi bir Y kromozomu mikrodelesyonu saptanmadı. Sonuç: Mevcut bilgiler ve geçmişteki literatür çalışmaları eşliğinde özellikle şiddetli oligospermi ve azospermili hastalarda kromozom analizi ve Y mikro delesyonu analizlerini yardımcı üreme tekniklerinden önce önermekteyiz.

Human umbilical cord matrix Wharton’s jelly mesenchymal stem cells (WJ-MSCs) are commonly utilized in regenerative medicine due to their therapeutic benefits. However, the microenvironmental stress present in patients with hyperglycemia can significantly reduce mesenchymal stem cell (MSC) viability under high-glucose conditions in the body, ultimately reducing their therapeutic effectiveness. Enhancing the survival rate of MSCs following cell transplantation remains a crucial challenge. This study investigates whether Quantum Hyperlight (QHL) can counteract the detrimental effects of high glucose (HG), thereby improving MSC survival, proliferation, and mitochondrial function. We aimed to evaluate the effect of QHL on cellular viability, proliferation, and mitochondrial activity in WJ-MSCs exposed to HG. MSCs were cultured in a medium containing normal glucose (NG) (1 g/L) and HG (4.5 g/L). MSCs in the HG medium were exposed to QHL for 90 s or 180 s with an energy density of 2.4 Joules/cm ² /minute and an average power density of 40 mW/cm ² . Then, proliferating cell nuclear antigen (PCNA), MTT assays, and Mitotracker Green staining were performed to evaluate cell viability and proliferation. The viability of MSCs was significantly increased in the QHL-treated groups (84% in QHL-90 s and 86% in QHL-180 s) compared to the untreated HG group (65%, p < 0.001). PCNA expression in QHL-90 s and QHL-180 s groups showed significant increases ( p < 0.001) compared to the untreated HG group. MitoTracker staining intensity was significantly higher in the QHL-treated groups compared to the untreated HG group ( p < 0.001). The HG environment reduced viability, proliferation, and mitochondrial staining. In the context of the NG environment, MSCs exhibited notable differences. However, the viability, proliferation, and mitochondrial staining rates of MSCs were significantly higher in the HG conditions when treated with QHL compared to the group that did not receive QHL. This study introduces QHL as a novel approach to enhance the therapeutic potential of WJ-MSCs under HG conditions, demonstrating its ability to improve cellular viability, proliferation, and mitochondrial activity. This study highlights its potential as a pretreatment to improve clinical outcomes in regenerative medicine.