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  3. Investigating the effect of lung and tail tissue extracts on the differentiation of mesenchymal stem cells into nerve cells

Investigating the effect of lung and tail tissue extracts on the differentiation of mesenchymal stem cells into nerve cells

Number of pages: 66 File Format: word File Code: 32096
Year: Not Specified University Degree: Not Specified Category: Research Methodology
Tags/Keywords: distinction - Extract of lung tissue - fetal tail - FGF - induction - mesenchymal - mitogen - Nerve cells - Neuroectoderm - Neuron cells - Six extract - Stem cells
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  • Summary of Investigating the effect of lung and tail tissue extracts on the differentiation of mesenchymal stem cells into nerve cells

    Abstract:

    Introduction and purpose: One of the most important adult stem cells that has attracted the attention of most researchers is mesenchymal stem cells. Mesenchymal stem cells derived from bone marrow have the ability to transform into different types of cells, including fat cells, bone and cartilage, in addition, these cells can be differentiated into neuronal cells. Most of the substances that have been used to induce stem cells into nerve cells such as: retinoic acid, dimethyl sulfoxide, deprenyl, etc. The compounds are toxic and expensive. In this research, fetal lung and tail tissue extracts were used as non-toxic inducers.

    Materials and methods: Mesenchymal stem cells were prepared and cultured from the femur bone of adult BALB/C mice. After 3 passages, they were induced by fetal lung and tail tissue extracts with concentrations of 50%, 70% and 80% in one group for 3 days and in the other group for 7 days. Then, they were examined by morphological and flow cytometry methods. In the morphological examination, cresyl violet was used and in the flow cytometry method, nestin and tubulin antibodies were used to evaluate the neural phenotype.

    Findings: The flow cytometry results showed that the expression of the nestin marker decreased significantly (P>0.05) in the cells after induction for 3 days compared to the control group, but after induction for 7 days, the expression of these two Markero also found a significant increase (P>0.05) in the expression of tubulin marker in induction for 3 days. Morphological changes were also in line with flow cytometry results.

    Conclusion: Treatment of mesenchymal stem cells with lung and tail tissue extracts increases the amount of neurons. This study showed that mesenchymal cells have the ability to differentiate into neurons in vitro and this issue depends on the type of induction method.

    Key words: mesenchymal stem cells, neuronal cells, induction, differentiation, fetal lung and tail extract

    Introduction:

    The concept of stem cells is obtained from the rules governing the development of the embryo. Developmental biology involves the study of the processes of the stem cell concept, which transforms the rules of the fertilized egg into the complex structure and function of the organs of a healthy child. In fact, a stem cell can be called a mother cell, that is, a cell that can be the generator of other cells.

    From the point of view of scientists, a stem cell is considered as an undifferentiated cell that reproduces and self-regenerates and produces different cell lineages. In this way, the fertilized egg is considered as an omnipotent stem cell because it has the ability to produce all types of cells and create a complete organism. During the process of morphogenesis, cells multiply, migrate and differentiate. Some of the cells make the membranes around the embryo and some others form the mass of cells inside the embryo. These cells have the power to form all tissues and cells. The internal mass of the embryo contains cells that have unlimited self-regeneration power and can transform into all 3 layers of the embryo, these cells are considered as multipotent cells to differentiate into other lineages based on the activated need in the future. These cells are called multipotent cells that have the ability to differentiate into many tissues. The most obvious example of these cells is mesenchymal stem cells. Multipotent cells are finally differentiated into unipotent or prerequisite stem cells that are only capable of producing one cell line and have limited self-renewal power. These cells can be found in adult tissues. One of the important characteristics of stem cells is the differentiation of these cells. These cells have the power to differentiate into bone, cartilage, muscle, fat cells, and can also differentiate into nerve cells. The use of stem cells differentiated into nerve cells in the repair of damage to the nervous system and the treatment of Alzheimer's and Parkinson's is one of the most important programs of medical science today. Since access to a source of suitable stem cells is an important principle in cell therapy and bone marrow is an available and suitable source of mature stem cells, many researches have been conducted in the field of using bone marrow stem cells.

    Chapter One

    1-1- Review of past research:

    Research on stem cells started around 1970 (14). There are many reports on the differentiation of neurons from mouse and human stem cells. Based on the studies of Bain and his colleagues in 1995, they showed the effect of retinoic acid and the effect of areas with rosette morphology on embryonic stem cells and then the differentiation of these cells into nerve cells. Also, in 2001, Zhang introduced areas with rosette morphology as primary neuroectoderm inducing areas (6). Okab and his colleagues have introduced FGF as a mitogenic factor in the proliferation of differentiated stem cells, and other researchers such as Rao, Li, and Mojtaba were able to obtain a greater number of differentiated neural cells in vitro with the changes they made in the simple Bain method (12, 55).  Rolletshek and his colleagues obtained 200-cell three-dimensional colonies of embryonic-like bodies [1] using the hanging drop method and cultured them in the form of a suspension in a serum-free medium, and after transferring them to a plate covered with polylysine and laminin in the presence of bFGF2 and EGF3, neural progenitor cells proliferated and formed dopaminergic neurons (59).

    Stem cells Located in the bone marrow, they were first identified by Friedenstein and Petrakova in 1966. In a study, these researchers extracted and described bone progenitor cells from rat bone marrow. But definitive evidence was presented by Friedenstein in 1970 (4). The researcher cultured the bone marrow samples in a plastic dish and discarded the non-adherent cells after 4 hours or so. Originally, these discarded cells were hematopoietic cells. He reported that a small part of the bone marrow cells was heterogeneous in appearance, in fact, the part that makes strong connections with the surface of the culture dish is sticky. These aggregations remained dormant for 2 to 4 days and then rapidly proliferated. These cells appeared uniformly spindle-shaped after several passages (4). Friedenstein's initial observation was developed by several researchers, especially Piersma and Owen et al. in 1980 (57). Previous studies have shown that bone marrow mesenchymal cells have the ability to differentiate into nerve cells (41). Various studies have been conducted in the field of differentiating these cells into neuronal cells.

    The first efforts in the field of differentiating MSCs into neuronal cells were started by Woodbury. He did this using mercaptoethanol and DMSO[2] on human MSCs and observed that although noticeable morphological changes occur in these cells, this induction is not an efficient method because the resulting cells They were functionally inactive. Anyway, this report was the first promise in the field of applying this type of stem cells in the treatment of spinal cord injuries. Further, by using the growth factors forskolin and bFGF and preparing a suitable substrate of protein L-lysine and concanavalin A and a combination of chemical inducers, he was able to differentiate rat mesenchymal stem cells into nerve cells by about 60%. He stated that the molecular mechanism of this differentiation is the turning off of the expression of mesenchymal genes in contrast to the start of the expression of neuronal genes. Also, Andreas and Herman were able to culture these cells in the form of a suspension of complex cells similar to neurospheric structures from mesenchymal cells using serum-free medium and growth factors bFGF and EGF. They found that MSCs normally express 3-4% nestin protein and after induction using growth factors on these progenitor cells, they did not express any of the specific proteins of mature neuronal cells (21, 31, 45, 67, 70). The results of Sanchez and Woodbury's research in 2000 confirmed the differentiation of mesenchymal cells into neural cells in the laboratory environment with the molecular western blot method (43). According to another report, in 2002, Jiang and his colleagues isolated a population of stem cells from the bone marrow of mice and injected them into adult mice. These cells were differentiated into blood cells and nerve cells (48).

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    5- Baghban Islamejad Mohammad, Nadri Samad, Isolation and proliferation of mouse mesenchymal stem cells: the effect of cell density on Morphology, differentiation and expression of cell surface markers, 13, 1, 1387, 49-37

    6- Baharond Hossein, Hatami Maryam, Zare Narges, the effect of retinoic acid on the differentiation of neural cells from human embryonic stem cells with rosette and non-rosette morphology, Journal of Cell Medicine, 6, 22, 1383, 109-103

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    11- Khayat Rodbari Nasim, Pariorkazem, Mohseni Kochsefhani Homa, Yaghmai Paricher, study of the effect of growth factors in adult liver extract on differentiation Fetal cord stem cells to blood cells in vitro, Journal of Medical Sciences of Islamic Azad University, 19, 1, 2018, 48-41

    12- Jahanbakht Hedah, Parivar Kazem, Akhundi Mohammad Mahdi, Jadi Tehrani Mahmoud, Modares Mohammad Hossein, Sadeghi Mohammad Reza, Karamtipour Mohammad, Zarnani Amir Hossein, Shida Saleh Kho, investigating the effect of growth factor, NGF, RA, Shh on the differentiation of mouse embryonic stem cells. to valigocytic nerve cells, Fertility and Infertility Quarterly, 1385, 357-349

    13- Mina Ramezani, Fatemeh Piriaei, Aini Fatemeh, Falah Reofi Mehri, Isolation, proliferation and differentiation of chicken bone marrow mesenchymal stem cells into bone, cartilage and fat cells, Islamic Azad University of Life Sciences Quarterly, Zanjan Branch, 7, 4, 1388, 44-35

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