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The Production of Pluripotent Stem Cells from Mouse Amniotic Fluid Cells Using a Transposon System
Published on: February 28, 2017
The human amniotic fluid mesenchymal stem cells therapy on, SKOV3, ovarian cancer cell line
Shiva Gholizadeh-Ghaleh Aziz1, Zahra Fardyazar2,3, Maryam Pashaiasl2,3,4,5
1Department of Biochemistry, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.
Purpose:
One of the most common malignancies peculiar to female health with few symptoms, low response to therapy, difficult diagnosis, frequent relapse, and high mortality, is ovarian cancer. Thus, our experiment, using Human amniotic fluid mesenchymal stem cells (hAFMSCs) as a therapeutic tool, aims to find an efficient treatment approach for patients suffering from SKOV3 ovarian cancer.
Material & Methods:
In this study, we obtained 5 ml amniotic fluid from 16-20 week pregnant women who underwent amniocentesis for routine prenatal diagnosis by karyotyping in Al-Zahra Hospital of Tabriz University of Medical Sciences, Iran. Using trans wells in 24 wells plate, hAFMSCs were isolated from all samples, co-cultured with SKOV3 ovarian cancer cell line, and characterized via flow cytometry and RT-PCR. Human skin fibroblast cells (HSFCs) were isolated and used as a negative control. SKOV3 and HSFCs' viability after 5 days was evaluated by MTT assay. Cell cycle and apoptotic genes were analyzed by real-time PCR.
Results:
We successfully isolated and characterized hAFMSCs through it positivity for CD44 and CD90 specific mesenchymal stem cell markers and negativity for CD31 and CD45. Oct4 and NANOG were evaluated by RT-PCR as pluripotency markers, and visualized on 2% gel electrophoresis. We established hAFMS cell lines after 5 days of co-culturing the SKOV3 cells, viability was decreased; however, HSFCs did not show toxicity by MTT assay. The genes indicated upregulation and high expression by a real-time PCR.
Conclusions:
Our findings showed that hAFMSCs have natural tumor tropism, and can release soluble factors in a cell culture, which cause an efficient anticancer effect. Thus, we can use hAFMSCs for complete anticancer therapy on SKOV3 cell line at cell culture condition and possibly in vivo in the near future.
Insights
Human amniotic fluid mesenchymal stem cells (hAFMSCs) show promise in treating ovarian cancer by targeting SKOV3 cells. These stem cells demonstrated natural tumor tropism and induced an anticancer effect in cell cultures.
Area of Science:
- Biomedical Science
- Stem Cell Research
- Oncology
Background:
- Ovarian cancer is a leading cause of cancer death in women, characterized by late diagnosis and poor treatment response.
- Developing novel therapeutic strategies is crucial for improving patient outcomes in ovarian cancer.
Purpose of the Study:
- To investigate the potential of human amniotic fluid mesenchymal stem cells (hAFMSCs) as a therapeutic agent against SKOV3 ovarian cancer cells.
- To evaluate the efficacy and safety of hAFMSCs in a cell culture model.
Main Methods:
- hAFMSCs were isolated from amniotic fluid and characterized using flow cytometry and RT-PCR.
- SKOV3 ovarian cancer cells were co-cultured with hAFMSCs, and cell viability was assessed using MTT assay.
- Gene expression related to cell cycle and apoptosis was analyzed by real-time PCR.
Main Results:
- hAFMSCs were successfully isolated and characterized, expressing key stem cell markers (CD44, CD90) and pluripotency markers (Oct4, NANOG).
- Co-culturing with hAFMSCs significantly reduced SKOV3 cell viability without causing toxicity to human skin fibroblast cells (HSFCs).
- Real-time PCR indicated upregulation of genes associated with anticancer effects.
Conclusions:
- hAFMSCs possess inherent tumor-homing abilities and secrete factors that exert potent anticancer effects on ovarian cancer cells.
- hAFMSCs hold significant potential for developing novel ovarian cancer therapies, with prospects for in vivo application.
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