scholarly journals Differentiation of bone marrow derived mesenchymal stem cells into male germ-like cells in co-culture with testicular cells

2019 ◽  
Vol 53 (2) ◽  
pp. 93-99
Author(s):  
Nasim Malekmohamadi ◽  
Alireza Abdanipour ◽  
Mehrdad Ghorbanlou ◽  
Saeed Shokri ◽  
Reza Shirazi ◽  
...  

AbstractObjective. Stem cell therapy, specifically, pre-induction of mesenchymal stem cells toward male germ-like cells may be useful in patients with azoospermia. The aim of this study was to evaluate in vitro differentiation of mouse bone marrow-derived mesenchymal stem cells (BMSCs) into male germ-like cells by indirect co-culture with testicular cells in the presence of bone morphogenetic protein 4 (BMP4).Methods. Experimental groups included: control (mouse BMSCs), treatment group-1 (BMSCs treated with BMP4), treatment group-2 (indirect co-culture of BMSCs with mouse testicular cells in the presence of BMP4) and treatment group-3 (indirect co-culture of BMSCs with testicular cells). BMSCs-derived male germ-like cells were evaluated by the expression of Dazl, and Stra8 using RT-qPCR.Results. Stra8 gene expression was significantly increased in the treatment group-2 and Dazl gene was significantly increased in the treatment group-1 compared to other groups. In conclusion, indirect co-culturing of BMSCs with testicular cells and BMP4 leads to the differentiation of BMSCs into male germ-like cells which express specific male germ-like genes. Testicular cells released factors that contributed to the differentiation of BMSCs into male germ progenitor cells.Conclusion. This study suggests that mesenchymal stem cells may be differentiated into male germ-like cells and therefore, may be a novel treatment option for men with azoospermia.

2016 ◽  
Vol 19 (2) ◽  
pp. 111-116
Author(s):  
Rafal Hussamildeen Abdullah ◽  
◽  
Shahlla Mahdi Salih ◽  
Nahi Yosef Yaseen ◽  
Ahmed Majeed Al-Shammari ◽  
...  

Biomedicines ◽  
2019 ◽  
Vol 7 (1) ◽  
pp. 7 ◽  
Author(s):  
Sara Mohamed ◽  
Shahinaz Shalaby ◽  
Soumia Brakta ◽  
Lelyand Elam ◽  
Amro Elsharoud ◽  
...  

Background: Premature ovarian insufficiency (POI) is a challenging disease, with limited treatment options at the moment. Umbilical cord blood mesenchymal stem cells (UCMSCs) have demonstrated promising regenerative abilities in several diseases including POI. Materials and Method: A pre-clinical murine case versus vehicle control randomized study. Two experiments ran in parallel in each of the three groups. The first was to prove the ability of UCMSCs in restoring ovarian functions. The second was to prove improved fertility. A total of 36 mice were randomly assigned; 6 mice into each of 3 groups for two experiments. Group 1 (control), group 2 (sham chemotherapy), group 3 (stem cells). Results: In the first experiment, post-UCMSCs treatment (group 3) showed signs of restored ovarian function in the form of increased ovarian weight and estrogen-dependent organs (liver, uterus), increased follicular number, and a significant decrease in FSH serum levels (p < 0.05) compared to group 2, and anti-Mullerian hormone (AMH) serum levels increased (p < 0.05) in group 3 versus group 2. Immuno-histochemistry analysis demonstrated a higher expression of AMH, follicle stimulating hormone receptor (FSHR) and Inhibin A in the growing follicles of group 3 versus group 2. In the second experiment, post-UCMSCs treatment (group 3) pregnancy rates were higher than group 2, however, they were still lower than group 1. Conclusion: We demonstrated the ability of UCMSCs to restore fertility in female cancer survivors with POI and as another source of stem cells with therapeutic potentials.


2011 ◽  
Vol 20 (9) ◽  
pp. 1549-1561 ◽  
Author(s):  
Olga Katsara ◽  
Louisa G. Mahaira ◽  
Eleni G. Iliopoulou ◽  
Ardiana Moustaki ◽  
Aristidis Antsaklis ◽  
...  

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Heng Zhang ◽  
Wen Zhang ◽  
Guangchao Bai ◽  
Lei Gao ◽  
Kuanxin Li

This study is aimed at investigating the effects of bone morphogenetic protein-7 (BMP-7) on the differentiation of bone marrow mesenchymal stem cells (BMSCs) into neuron-like cells in vitro. The rat BMSCs were isolated and identified, which were divided into the control, empty, recombinant rhBMP-7 transfection, and Lv-BMP-7 transfection groups. BMSCs were induced under different conditions. CCK-8 assay was performed to detect cell proliferation. ALP was used to detect cell activity. Cellular morphology after induction was observed. Immunofluorescence was conducted to detect the expression and location of nerve cell markers. Quantitative real-time PCR and Western blot analysis were performed to detect the mRNA and protein expression levels, respectively. The rhBMP-7 and Lv-BMP-7 promoted the proliferation of BMSCs, accompanied with increased ALP activities. Morphological observations revealed that rhBMP-7 and Lv-BMP-7 induced BMSCs to differentiate into neuron-like cells. Immunofluorescence revealed that the rhBMP-7 and Lv-BMP-7 groups showed positive expression of MAP-2 and Nfh in BMSCs. MAP-2 was mainly distributed in the cell body and cellular protrusion, while Nfh was mainly distributed in the cytoplasm and cell protrusion. Positive mRNA and protein expressions of MAP-2 and Nfh were observed in the cells of the rhBMP-7 and Lv-BMP-7 groups, and the expression levels were significantly higher than the control and empty groups. Both exogenous BMP-7 (rhBMP-7) and endogenous BMP-7 (Lv-BMP-7) can induce BMSCs to differentiate into neuron-like cells highly expressing the neuronal markers MAP-2 and Nfh.


2020 ◽  
Vol 66 (3) ◽  
pp. 265-273
Author(s):  
O.V. Vysotskaya ◽  
A.I. Glukhov ◽  
Yu.P. Semochkina ◽  
S.A. Gordeev ◽  
E.Yu. Moskaleva

In proliferating normal and tumor cells, the telomere length (TL) is maintained by high telomerase activity (TA). In the absence of TA the TL maintenance involves a mechanism of alternative lengthening of telomeres (ALT). The aim of this study was to investigate the level of TA, the mTert expression and TL in cultured normal and transformed by γ- and γ,n-irradiation mesenchymal stem cells (MSCs) from mouse bone marrow, in sarcomas that developed after the transplantation of these cells into syngeneic mice, and in fibrosarcoma cell lines obtained from these tumors to find out the role of AT or ALT in maintaining TL in these cells. During prolonged cultivation of normal and transformed under the influence of γ- (1 Gy and 6 Gy) and γ,n-irradiation (0.05 Gy, 0.5 Gy, and 2 Gy) MSCs from mouse bone marrow, a decrease in TA was detected in irradiated cells. Even deeper decrease in TA was found in sarcomas developed after administration of transformed MSCs to syngeneic mice and in fibrosarcoma cell lines isolated from these tumors in which TA was either absent or was found to be at a very low level. TL in three of the four lines obtained was halved compared to the initial MSCs. With absent or low TA and reduced TL, the cells of all the obtained fibrosarcoma lines successfully proliferated without signs of a change in survival. The mechanism of telomere maintainance in fibrosarcoma cell lines in the absence of TA needs further investigation and it can be assumed that it is associated with the use of the ALT. The detected decrease or absence of TA in transformed under the action of irradiation MSCs with the preservation or even an increase in the telomerase gene expression may be associated with the formation of inactive splicing variants, and requires further study. The obtained lines of transformed MSCs and fibrosarcomas with TA and without the activity of this enzyme can be a useful model for studying the efficacy of TA and ALT inhibitors in vitro and in vivo.


Author(s):  
Nugroho Setyowardoyo

The state of low oxygen levels known as hypoxia in humans is considered a dangerous condition is apparently a normal physiological condition and required by the stem cells as they are in the body. Mesencyimal Stem Cells (MSCs) require physiologically optimal conditions of low O2 tension of 1-3% in the bone marrow. The purpose of this study was to reveal the difference between in vitro culture of MSC in normoxia condition (20% O 2 concentration) with hypoxia condition (1% O2 content) especially in terms of viability, pluripotent properties, and MSC proliferation ability of the culture it produces. This research is an explorative laboratory research invitro on Bone Marrow Mesenchymal Stem Cells (BMSCs) culture using hypoxia condition. The study design used Randomized Control Group Post-Test Only Design. This research was conducted for 2 months. There was a significant difference in mean slow proliferation based on the number of Least-like CFU-Cs between the control group, treatment group 1 and treatment group 2, the mean percentage of the number of cells expressing the OCT4 coding gene on immunofluorosense examination between the control group, 1 and the treatment group 2 and the mean percentage of cell numbers expressing the OCT4 coding gene on the immunofluorosense examination between the control group, the treatment group 1 and the treatment group 2 showing p <0.01. There was a significant difference of percentage of non-absorbing color cell number of trypsin blue (viable cells) between control group, treatment group 1, and treatment group 2 showing p value <0.05. This suggests that the precondition of culture with normoxia provides an opportunity for cells to adapt and proliferate before being conditioned in hypoxic cultures. Cultures with hypoxic conditions and preconditions of normoxia are the best culture conditions because they produce cells that are capable of maintaining pluripotency properties while still having better proliferation and viability capability compared with direct hypoxia conditions.Keywords: Hypoxia, normoxia, bone marrow, mesenchymal stem cells.


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