Showing 9 results for Stromal Cells
Mehri Fayazi, Mojdeh Salehnia, Saeideh Ziaei,
Volume 14, Issue 7 (7-2016)
Abstract
Background: Stem cell factor (SCF) is a transcriptional factor which plays crucial roles in normal proliferation, differentiation and survival in a range of stem cells.
Objective: The aim of the present study was to examine the proliferation effect of different concentrations of SCF on expansion of human endometrial CD146+ cells.
Materials and Methods: In this experimental study, total populations of isolated human endometrial suspensions after fourth passage were isolated by magnetic activated cell sorting (MACS) into CD146+ cells. Human endometrial CD146+ cells were karyotyped and tested for the effect of SCF on proliferation of CD146+ cells, then different concentrations of 0, 12.5, 25, 50 and 100 ng/ml was carried out and mitogens-stimulated endometrial CD146+ cells proliferation was assessed by MTT assay.
Results: Chromosomal analysis showed a normal metaphase spread and 46XX karyotype. The proliferation rate of endometrial CD146P + P cells in the presence of 0, 12.5, 25, 50 and 100 ng/ml SCF were 0.945±0.094, 0.962±0.151, 0.988±0.028, 1.679±0.012 and 1.129±0.145 respectively. There was a significant increase in stem/ stromal cell proliferation following in vitro treatment by 50 ng/ml than other concentrations of SCF (p=0.01).
Conclusion: The present study suggests that SCF could have effect on the proliferation and cell survival of human endometrial CD146P+P cells and it has important implications for medical sciences and cell therapies.
Azadeh Esmaeli, Mojgan Moshrefi, Ali Shamsara, Seyed Hasan Eftekhar-Vaghefi, Seyed Noureddin Nematollahi-Mahani,
Volume 14, Issue 9 (9-2016)
Abstract
Background: Fetal bovine serum (FBS) is widely used in cell culture laboratories,risk of zoonotic infections and allergic side effects create obstacles for its use inclinical trials. Therefore, an alternative supplement with proper inherent growthpromotingactivities is demanded.
Objective: To find FBS substitute, we tested human umbilical cord blood serum(hUCS) for proliferation of human umbilical cord matrix derived mesenchymal stemcells (hUC-MSCs) and human bone marrow-derived mesenchymal cells (hBMMSCs).
Materials and Methods: Umbilical cord blood of healthy neonates, delivered byCaesarian section, was collected and the serum was separated. hUC-MSCs andhBM-MSCs were isolated and characterized by assessment of cell surface antigensby flow cytometry, alkaline phosphatase activity and osteogenic/adipogenicdifferentiation potential. The cells were then cultured in Iscove's ModifiedDulbecco's Medium (IMDM) by conventional methods in three preparations: 1- withhUCS, 2- with FBS, and 3- without serum supplements. Cell proliferation wasmeasured using WST-1 assay, and cell viability was assessed by trypan bluestaining.
Results: The cells cultured in hUCS and FBS exhibited similar morphology andmesenchymal stem cells properties. WST-1 proliferation assay data showed nosignificant difference between the proliferation rate of either cells following hUCSand FBS supplementation. Trypan blue exclusion dye test also revealed nosignificant difference for viability between hUCS and FBS groups. A significantdifference was detected between the proliferation rate of stem cells cultured inserum-supplemented medium compared with serum-free medium.
Conclusion: Our results indicate that human umbilical cord serum can effectivelysupport proliferation of hBM-MSCS and hUC-MSCs in vitro and can be used as anappropriate substitute for FBS, especially in clinical studies.
Mina Jafarabadi, Mojdeh Salehnia, Rana Sadafi,
Volume 15, Issue 1 (1-2017)
Abstract
Background: The animal models of endometriosis could be a valuable alternative tool for clarifying the etiology of endometriosis.
Objective: In this study two endometriosis models at the morphological and molecular levels was evaluated and compared.
Materials and Methods: The human endometrial tissues were cut into small fragments then they were randomly considered for transplantation into γ irradiated mice as model A; or they were isolated and cultured up to fourth passages. 2×106 cultured stromal cells were transplanted into γ irradiated mice subcutaneously as model B. twenty days later the ectopic tissues in both models were studied morphologically by Periodic acid-Schiff and hematoxylin and eosin staining. The expression of osteopontin (OPN) and matrix metalloproteinase 2 (MMP2) genes were also assessed using real time RT-PCR. 17-β estradiol levels of mice sera were compared before and after transplantation.
Results: The endometrial like glands and stromal cells were formed in the implanted subcutaneous tissue of both endometriosis models. The gland sections per cubic millimeter, the expression of OPN and MMP2 genes and the level of 17-β estradiol were higher in model B than model A (p=0.03).
Conclusion: Our observation demonstrated that endometrial mesenchymal stromal cells showed more efficiency to establish endometriosis model than human endometrial tissue fragments
Mojdeh Salehnia, Mehri Fayazi, Shokreya Ehsani,
Volume 15, Issue 4 (6-2017)
Abstract
Background: Concerning the low population of human endometrial mesenchymal cells within the tissue and their potential application in the clinic and tissue engineering, some researches have been focused on their in vitro expansion.
Objective: The aim of this study was to evaluate the effect of leukemia inhibitory factor (LIF) as a proliferative factor on the expansion and proliferation of human endometrial stromal cells.
Materials and Methods: In this experimental study, the isolated and cultured human endometrial stromal cells from women at ovulatory phase aged 20-35 years, after fourth passage were divided into control and LIF-treated groups. In the experimental group, the endometrial cells were treated by 10 ng/ml LIF in culture media and the cultured cells without adding LIF considered as control group. Both groups were evaluated and compared for proliferation rate using MTT assay, for CD90 marker by flow cytometric analysis and for the expression of Oct4, Nanog, PCNA and LIFr genes using real-time RT-PCR.
Results: The proliferation rate of control and LIF-treated groups were 1.17±0.17and 1.61±0.06 respectively and there was a significant increase in endometrialstromal cell proliferation following in vitro treatment by LIF compared to controlgroup (p=0.049). The rate of CD90 positive cells was significantly increased in LIFtreatedgroup (98.96±0.37%) compared to control group (94.26±0.08%) (p=0.0498).Also, the expression ratio of all studied genes was significantly increased in the LIFtreatedgroup compared to control group (p=0.0479).
Conclusion: The present study showed that LIF has a great impact on proliferation, survival, and maintenance of pluripotency of human endometrial stromal cells and it could be applicable in cell therapies.
Athar Talebi, Nasim Hayati Roodbari, Hamid Reza Sameni, Sam Zarbakhsh,
Volume 18, Issue 7 (7-2020)
Abstract
Background: Apigenin is a plant-derived flavonoid with antioxidative and antiapoptotic effects. Bone marrow stromal cells (BMSCs) are a type of mesenchymal stem cells (MSCs) that may recover damaged ovaries. It seems that apigenin may promote the differentiation of MSCs.
Objective: The aim of this study was to investigate the effect of coadministration of apigenin and BMSCs on the function, structure, and apoptosis of the damaged ovaries after creating a chemotherapy model with cyclophosphamide in rat.
Materials and Methods: For chemotherapy induction and ovary destruction, cyclophosphamide was injected intraperitoneally to 40 female Wistar rats (weighing 180–200 gr, 10 wk old) for 14 days. Then, the rats were randomly divided into four groups (n = 10/each): control, apigenin, BMSCs and coadministration of apigenin and BMSCs. Injection of apigenin was performed intraperitoneally and BMSC transplantation was performed locally in the ovaries. The level of anti-mullerian hormone serum by ELISA kit, the number of oocytes by superovulation, the number of ovarian follicles in different stages by H&E staining, and the expression of ovarian Bcl-2 and Bax proteins by western blot were assessed after four wk.
Results: The results of serum anti-mullerian hormone level, number of oocytes and follicles, and Bcl-2/Bax expression ratio showed that coadministration of apigenin and BMSCs significantly recovered the ovarian function, structure, and apoptosis compared to the control, BMSC, and apigenin groups (p < 0.001).
Conclusion: The results suggest that the effect of coadministration of apigenin and BMSCs is maybe more effective than the effect of their administrations individually on the recovery of damaged ovaries following the chemotherapy with cyclophosphamide in rats.
Fatemeh Akyash, Mahdieh Javidpou, Ehsan Farashahi Yazd, Jalal Golzadeh, Fatemeh Hajizadeh-Tafti, Reza Aflatoonian, Behrouz Aflatoonian,
Volume 18, Issue 11 (11-2020)
Abstract
Background: Human endometrium with consecutive regeneration capability undergoes monthly hormonal changes for probable implantation, which confirms the presence of the cells in the basalis layer known as stem cell.
Objective: Previously, we reported the isolation and culture of the mesenchymal-like cells from human endometrium. In this study, we evaluated the biological and stemness characteristics of these cells.
Materials and Methods: The characterization of Yazd human endometrial-derived mesenchymal stem/stromal cells (YhEnMSCs) was assessed using immunofluorescence (IF) staining for CD105, VIMENTIN, and FIBRONECTIN as markers and RT-PCR for CD166, CD10, CD105, VIMENTIN, FIBRONECTIN, MHCI, CD14, and MHCII genes. Flow cytometry (FACS) was performed for CD44, CD73, CD90, and CD105 markers. Moreover, the differentiation capacity of the YhEnMSCs to the osteoblast and adipocytes was confirmed by Alizarin Red and Oil Red staining.
Results: YhEnMSCs expressed CD105, VIMENTIN, FIBRONECTIN, CD44, CD73, and CD90 markers and CD166, CD10, CD105, VIMENTIN, FIBRONECTIN, and MHCI, but, did not express CD14, MHCII.
Conclusion: Our data confirm previous reports by other groups indicating the application of endometrial cells as an available source of MSCs with self-renewal and differentiation capacity. Accordingly, YhEnMSCs can be used as a suitable source for cell-based therapies.
Shohreh Nikoo, Massoumeh Ebtekar, Mahmood Jeddi-Tehrani, Mahmood Bozorgmehr, Amir Hassan Zarnani,
Volume 19, Issue 1 (1-2021)
Abstract
Background: Menstrual blood-derived stromal/stem cells (MenSCs) are a new population of refreshing and highly proliferative stem cells. Immunomodulatory effects of MenSCs profoundly depend on their relative density.
Objective: To find whether MenSCs cultured at varying numbers would differentially affect the allogenic peripheral blood mononuclear cells (PBMCs) key features.
Materials and Methods: PBMCs were co-cultured with various MenSCs numbers. PBMCs proliferation was investigated via 3H-thymidine incorporation. Flow cytometry was used to assess human leukocyte antigen (HLA)-DR, HLA-ABC, HLA-G, and co-stimulatory markers on MenSCs and the percentage of regulatory T cells (Tregs) among PBMCs. The concentration of cytokines was determined in supernatant of co-cultures.
Results: The support of PBMCs proliferation at low MenSCs densities correlated with higher levels of pro-inflammatory interferon gamma (IFN-γ) in MenSCs/PBMCs co-culture and increased expression of HLA-DR by MenSCs. On the other hand, the suppressive property of MenSCs at higher densities was independent of Treg frequency, but correlated with a high concentration of Interleukin (IL)-6 and IL-10 in the co-cultures.
Conclusion: Totally, at different seeding densities, MenSCs could differentially interact with PBMCs leading to significant changes in the level of anti- and/or pro-inflammatory factors. These preliminary in vitro results are suggested to be taken into consideration in experimental models of MenSC-based immunomodulation. Nonetheless, for efficient utilization of MenSCs anti-inflammatory features in pre-clinical disease models, we still need to broaden our knowledge on MenSC-immune system cross-talk; this could play a part in designing more optimized MenSCs injection modalities in the case of future pre-clinical and subsequently clinical settings.
Zarbakhsh S, Safari R, Sameni Hr, Yousefi B, Safari M, Khanmohammadi N, Hayat P,
Volume 19, Issue 5 (5-2021)
Abstract
Background: Despite the great benefits of chemotherapy in treating cancer patients, it has some side effects on ovaries. Cyclophosphamide is one of the most used chemotherapy drugs which directly damages ovaries. It has been observed that transplantation of bone marrow stromal cells (BMSCs), a type of mesenchymal stem cells, may treat ovarian damage after chemotherapy. On the other hand, L-carnitine (LC), as a flavonoid antioxidant, appears to play an essential role in fatty acid metabolism and has beneficial effects on damaged ovaries. In addition, LC has beneficial effects on differentiation and reduction of apoptosis in BMSCs.
Objective: The aim of this study was to investigate the effects of co-administration of BMSC + LC on ovarian function, structure and apoptosis after creating a chemotherapy model with cyclophosphamide in rat.
Materials and Methods: Forty female Wistar rats were intraperitoneally injected with cyclophosphamide for 14 days for chemotherapy-induced ovarian destruction. Then, the rats were randomly divided into four groups: I. control group, 25 μl of culture medium was directly injected into the bilateral ovaries, II. BMSC group, 2×106 BMSCs suspended in 25 μl of culture medium were directly injected into the bilateral ovaries, III. LC group, 200 mg/kg of LC was injected intraperitoneally one day before until seven days after chemotherapy, IV. Co-administration of BMSC + LC group, injection of BMSCs, and LC were performed together. Four weeks later, the function of the ovaries was evaluated by measuring the levels of serum estradiol (E2) and follicle-stimulating hormone using the enzyme-linked immunosorbent assay kit, the structure of the ovaries was evaluated by counting the number of ovarian follicles at different stages using hematoxylin and eosin staining, and apoptosis was investigated by evaluating the expression of ovarian B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) proteins using western blot were assessed.
Results: Co-administration of BMSC + LC was more effective in repairing damaged ovaries than the effect of their separate administration. Co-administration of BMSC + LC increased E2 and decreased follicle-stimulating hormone levels compared to the control group (p < 0.001). The number of follicles was higher in the co-administration of BMSC + LC group compared to the control group (p < 0.001). Co-administration of BMSC + LC increased Bcl-2 protein level, decreased Bax protein level and increased Bcl-2/Bax ratio (p < 0.001).
Conclusion: The effect of co-administration of BMSC + LC is probably more effective than the effect of their separate administration on the recovery of damaged ovaries by chemotherapy with cyclophosphamide in rat.
Fatemeh Hajizadeh-Tafti, Jalal Golzadeh, Ehsan Farashahi-Yazd, Hassan Heidarian-Meimandi, Behrouz Aflatoonian,
Volume 20, Issue 7 (7-2022)
Abstract
Background: Fibroblasts from different parts of the human body have been used in cell biology, drug discovery and cell therapy studies. One of the most available sources of human fibroblasts is neonatal foreskin. Not only do these cells have wound-healing applications, but they are also the most popular source for pluripotent stem cell biotechnology. Moreover, several studies have indicated that different sources of fibroblasts display similar features to mesenchymal stem cells.
Objective: Generation and establishment of new human foreskin fibroblast cell lines called Yazd human foreskin fibroblasts (YhFFs).
Materials and Methods: In this lab resources study, the production of 3 YhFF cell lines (YhFF#8, YhFF#17, and YhFF#18) is reported. Their biological features were characterized using immunofluorescence, polymerase chain reaction, and flow-cytometry for mesenchymal markers such as fibronectin, vimentin, CD44, CD73, CD90, CD105, and hematopoietic markers CD34 and CD45.
Results: The YhFF cell lines were passaged more than 40 times and their normal karyotype was checked using G-binding. Similarly to previous reports, the flow cytometry analysis revealed that the YhFF cell lines displayed mesenchymal stromal cell characteristics.
Conclusion: This study will contribute to the development of clinical-grade cell-based products such as micro-vesicles and exosomes for future therapeutic applications in regenerative medicine.
