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Showing 4 results for Cell Differentiation
M. Rasekhi , B. Bakhshande , M. Sadeghizadeh , A. Salimi , M. Soleimani ,
Volume 9, Issue 1 (1-2018)
Abstract
Aims: The induction of artificial over-expression of miRNAs is an appropriate approach to more effective cell differentiation. The significant role of microRNA-1(miR-1) has been reported in the development and differentiation of cardiac cells. Lentivirus is an effective vector for stable cell line production. The aim of this study was the production of recombinant HEK293T with miR-1 overexpression as a biological model for cardiac studies.
Materials & Methods: In this experimental study, HEK 293T cells were cultured in DMEM medium with 10% Fetal Bovine Serum (FBS) and L-glutamine 2mM and Penicillin-Streptomycin 1X in incubator medium. After cloning of miR-1 gene, recombinant clones were selected and the recombination was confirmed by sequencing. The miR-1 carrying vector and auxiliary vectors were packaged in the HEK293T to produce the recombinant virus. The infection of HEK293T by recombinant virus was performed in order to achieve stable cell line. Then, GFP fluorescent marker evaluated the efficiency of transfection and effective virus dilution. Finally, the alteration in expression level of miR-1 was assessed by qPCR. Data analysis was performed by comparing the threshold cycle and Pfaffl method.
Findings: The most GFP expression was detected in transfected cells by 150 micromole dilution. GFP fluorescent marker facilitated optimization and purification of recombinant cells. qPCR investigation demonstrated the significant increase in expression of miR-1 in transfected cells in comparison to controls.
Conclusion: The stable recombinant HEK293T miR-1 over-expressing cell line in lentivirus can be utilized as a suitable biological model for investigation of cardiac evolution and development processes.
Bahram Ahmadian, Bahman Vahidi,
Volume 13, Issue 4 (1-2023)
Abstract
Evaluating the response of the stem cells to different mechanical stimulation is an important issue to obtain control over cell behavior in the culture environment. One of the effective parameters in the mechanoregulation of stem cells is the microstructure of scaffolds. Evaluating the effect of microstructure of scaffold in the lab environment is very complicated. Therefore, in this study, the effect of scaffold architecture on mechanical factors in the scaffold was investigated under oscillatory fluid flow by using numerical modeling. In this study, distribution of shear stress and fluid velocity in three types of scaffolds with spherical, cubical and regular hexagonal pores with length of 300, 350, 400, 450 and 500 micrometers were investigated by using computational fluid dynamics method. The results of the computational fluid dynamics model showed that the scaffold with spherical and cubic pores shape with length of 500 micrometers and scaffold with hexagonal pores with length of 450 micrometers experienced shear stress in the range of 0.1-10 mPa. This range of the shear stress is suitable for differentiation of the stem cell to bone cells. Moreover, the result of exerting oscillatory fluid flow to these scaffolds indicated that dead zones of the scaffold, where isn’t suitable for cell seeding, was decreased due to the access of fluid flow to the different area of scaffold. The results of this study can be used in a laboratory to achieve optimal stem cell culture to provide suitable environment culture for differentiation of stem cells toward the bone cell.
Volume 22, Issue 1 (12-2018)
Abstract
Primordial germ cells (PGCs) are the specialized cells that are created from epiblast cells and after the migration differentiate into spermatogonial cells. Also, Spermatogonial cells differentiate into spermatids during the spermatogenesis process. Created disorders in each of these stages cause infertility, so the recognizing of the mechanism of these cells from the early stages of formation to the differentiation and investigating the effective factors in differentiation can be useful in the treatment of the infertile people. Today, the cultivation of spermatogonial cells and transplantation of these cells can be effective in the investigation of spermatogonial stem cell and the treatment of infertility. In this paper, the formation and migration of primordial germ cells, the spermatogenesis process and the effective factors in differentiation of spermatogonial stem cells are investigated.
Volume 22, Issue 1 (12-2018)
Abstract
Introduction: Due to increase of infertile couples, potential differentiation and proliferation of umbilical cord mesenchymal stem cells (MSCs) and bone marrow stem cells (BM-MSCs) was compared to find proper stem cells for differentiation into germ-like cells.
Materials & Methods: In this experimental study, isolated umbilical cord and bone marrow mesenchymal stem cells were treated by Retinoic acid (10-6M) and Sertoli cells condition medium. Viability percentage and the rate of proliferation (population doubling time) of cells was calculated in both groups. The number of colonies was evaluated in different days of culture, and finally the expression of and meiotic genes investigated by RT-PCR.
Findings: The viability percentage was higher in BM-MSCs group and the rate of proliferation of cells increased by elevating the passage number. The number of colonies in the bone marrow stem cells was significantly higher than that of the umbilical cord MSCs (p<0.05). In contrast, the expression of PLZF, OCT4 and SCP3 genes were detected in umbilical cord MSCs after 10 days of culture. However, in BM-MSC, the expression of PLZF and SCP3 genes was observed only after 15 days of culture.
Conclusion: It seems that the human umbilical MSCs higher differentiation potential for producing germ-like cells when compared to the Bone marrow stem cells. In contrast, the proliferation potential of BM-MSCs is greater than umbilical cord MSCs. This difference is probably due to secreted growth factors from these cells.
