Objective: Adipose derived stem cells (ASCs) secrete numerous neurotrophic factors and cytokines in conditioned medium (CM), which protect neurons by its antioxidative and trophic effects. This research assesses the neuroprotective effect of ASCCM on neurotrophins genes expressions and tyrosine hydroxylase positive (TH+) cell density in male Wistar rats lesioned by 6-hydroxydopamine (6-OHDA).Materials and Methods: In this experimental study, the groups consisted of lesioned and sham rats with unilateral injections of 20 μg of 6-OHDA neurotoxin and phosphate buffered saline (PBS) into the striatum, respectively. Another groups received intravenous injections of 3×106 cells (ASCs group), 500 ml of CM (ASC-CM group) or medium [α-minimal essential medium (a-MEM) group)]. All rats underwent evaluations with the rotarod and apomorphine-induced rotation tests at 2, 4, 6, and 8 weeks post-injection. At 8 weeks we sacrificed some of the animals for real-time polymerase chain reaction (PCR) analysis, and evaluation of TH+cell counts.Results: We observed a significant decrease in contralateral turns to the lesions in the ASCs and ASC-CM groups compared to the neurotoxin lesioned or a-MEM groups at 8 weeks post transplantation. Cell and CM- injected rats showed a significant increase of staying on the rotarod compared to the lesion or a-MEM groups. Cell and CM-treated rats showed significant increases in theNGF and NT3 genes, respectively, compared with the lesion group. Both treated groups showed significant increases inBDNF gene expression and TH+cell density.Conclusion: The results suggested that ASCs and ASC-CM protected dopaminergic neurons through the expressions of neurotrophin genes.

Objective: In vivo, growth factors that mediate local cellcell interactions are responsible for the differentiation of primordial germ cells (PGCs) into germ stem cells after the arrival at the genital ridge. in vitro differentiation of PGCs is also dependent on their sequential exposure and response to an array of growth factors. In the presence of multiple growth signals, PGCs restart rapid proliferationin vitro and transform into pluripotent embryonic germ cells (EGCs). Hence, some cases of sterility are due to lack of germ cells, in the future, this problem could be treated using stem cell therapy. Thus, in this study we effort the preparation of testicular cell conditioned medium from rat testis.Materials and Methods: Testes of 1-day-old newborn male rats were removed and placed into tripsin EDTA solution. Then the harvested cells cultured with DMEM medium supplemented with 10% FCS, 1% nonessential amino acids, and 1% penicillin/streptomycin. After proliferation of germ cells, testiculat conditioned medium was collected every 3 days starting 10 days after initiation.Results: An obvious germ cell proliferation was evident in TCCs 8-10 days from initiation. The round cells were clustered together and easily dissociated from other cells when dishes were shaken. Germ cells differed in size and appeared as single cells or attached to each other, forming pairs or rows.Conclusion: Testis, particularly of newborns, contains germ cells that release growth factors during in vivo culture. In this regard, we try the in vivo differentiation of mesenchymal stem cells of rats into PGCs and precursors of sperm by TCC medium induction. Their results will be announced later.

Background: To evaluate the efficiency of Menstrual blood Stromal/Stem Cells (MenSCs) administration in Myocardial Infarction (MI), the effects of MenSCs and their derived conditioned Medium (CM) on cardiac function in MI rat model was assessed. Methods: Animals were divided into four groups including sham group, MI group, MenSCs derived CM group (CM group), and MenSCs suspended in CM (MenSCs+ CM) group. The injection of different groups was carried out 30 min after ligation of left anterior descending coronary artery into the infarct border zone. Results: The results showed a significant reduction in scar size after injection of MenSCs+CM compared to MI group. Ejection fraction and fractional shortening of MenSCs+CM group were higher than CM and MI group at day 28. Administration of MenSCs+CM led to much more survival of cardiomyocytes, and prevention of metaplastic development. Moreover, human mitochondrial transfer from MenSCs to cardiomyocytes was seen in group treated by MenSCs+CM. Indeed, MenSCs+CM treatment evoked nuclear factor-κ, B (NF-κ, B) down-regulation more than other treatments. Conclusion: MenSCs+CM treatment could significantly ameliorate cardiac function by different mechanisms including inhibition of cartilaginous metaplasia, inhibition of NF-κ, B and mitochondrial transfer.

Dear Editor, Mesenchymal stem cells (MSCs) are an appealing candidate for tissue regeneration due to their ability to differentiate into many cell types (1). However, the use of stem cells has several problems such as complicated safety and quality management and high cost and difficult cell handling (2). On the other hand, MSCs secrete various trophic factors such as cytokines and growth factors that may regenerate damaged tissues (3). Due to limited survival and differentiation of transplanted MSCs, it is proposed that paracrine effect of MSC secretion is the principal mechanism for tissue regeneration…

Background: This study aimed to investigate the maturation and fertilization rates of immature mouse oocytes using Embryonic Stem Cell Conditioned Medium (ESCM).Methods: Germinal Vesicle (GV) stage oocytes were observed in 120 NMRI mice, aged 4-6 weeks. GV oocytes with or without cumulus cells were subjected to IVM in either ESCM, Embryonic Stem Cell Growth Medium (ESGM), or a-minimum essential medium (a-MEM). After recording the Metaphase II (MII) oocyte maturation rate, the oocytes were fertilized in vitro. The fertilization success rate was recorded after 24 hr. The embryos were maintained in potassium Simplex Optimization Medium (KSOM) for 96 hr and allowed to grow until the blastocyst stage. After recording developmental competence, they were transferred into the uteri of pseudopregnant mice and their birth rates were recorded.Results: No significant difference existed between the maturation rates in a-MEM (68.18%) and ESCM (64.67%; p>0.05), whereas this rate was significantly higher for both a-MEM and ESCM compared to ESGM (32.22%; p<0.05). A significant difference in IVF success rate existed for oocytes grown in α-MEM (69.44%), ESCM (61.53%), and ESGM (0%). A significantly higher developmental competence was observed at the blastocyst stage for oocytes grown in a-MEM (51.2%) compared to ESCM (35%; p<0.05). 17 days after embryo transfer into the uteri of pseudopregnant mice, there was a nonsignficant (p>0.05), similar birth rate between α-MEM and ESCM (47 vs. 40%).Conclusion: ESCM is an effective medium for preantral follicle growth, oocyte maturation, and subsequent embryo development.

Purpose: Macrophages with an anti-inflammatory phenotype are critical for resolving inflammation and preventing chronic tissue injury. Estradiol is known to promote this favorable macrophage profile. This study evaluated the therapeutic potential of the secretome, delivered as conditioned medium, from estradiol-treated macrophages in experimental rheumatoid arthritis (RA) in Wistar rats. Methods: Rheumatoid arthritis was induced in Wistar rats using complete Freund’s adjuvant. Animals were assigned to five groups: healthy controls, arthritic rats receiving vehicle, arthritic rats treated with prednisolone, arthritic rats treated with conditioned medium from untreated macrophages, and arthritic rats treated with conditioned medium from estradiol-exposed macrophages. The lyophilized media were administered intraperitoneally on days 4, 12, and 20 post-induction,the study ended on day 24. Results: Conditioned medium from estradiol-treated macrophages exhibited significantly higher levels of anti-inflammatory mediators such as interleukin-10 (IL-10), transforming growth factor-beta, and indoleamine 2, 3-dioxygenase, along with increased messenger RNA expression of regulatory genes including early growth response 2 and mannose receptor. In vivo, this treatment notably reduced arthritis severity and improved weight gain compared to medium from untreated macrophages. These effects correlated with a marked decrease in antigen-specific proliferation and serum levels of inflammatory markers such as C-reactive protein (CRP), myeloperoxidase (MPO), nitric oxide (NO), IL-1, and tumor necrosis factor-alpha. Additionally, bone-destructive factors like receptor activator of nuclear factor kappa-B ligand (RANKL) and matrix metalloproteinase-9 (MMP-9) were significantly downregulated in treated rats. Conclusion: The conditioned medium derived from estradiol-treated macrophages, enriched with anti-inflammatory and regulatory components, presents a promising cell-free therapeutic strategy for immunotherapy in RA.