Development
Faeze Zarean; Somayeh Arabzadeh; Sarah Rajabi; Saeideh Erfanian; Mahmood Talkhabi
Abstract
Bone is the hardest and one of the most important tissues in the body. In case of bone damage, the current treatments do not completely repair and regenerate the bone. For this reason, cell-based tissue engineering strategies, especially Mesenchymal Stem Cells (MSCs), have received attention. MSCs have ...
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Bone is the hardest and one of the most important tissues in the body. In case of bone damage, the current treatments do not completely repair and regenerate the bone. For this reason, cell-based tissue engineering strategies, especially Mesenchymal Stem Cells (MSCs), have received attention. MSCs have the ability to self-renew and differentiate into different cell types, including bone cells, cartilage cells, and fat cells, among others. They are found in various tissues throughout the body, including bone marrow, adipose tissue, and umbilical cord tissue. Today, MSCs are a valuable resource for regenerative medicine and tissue engineering applications. In addition to the cells, scaffolds are another essential element of tissue engineering. One of these scaffolds is decellularized tissue-derived hydrogels, which are three-dimensional network of hydrophilic polymer chains that can absorb and retain a significant amount of water. In tissue engineering, they mimic the natural extracellular matrix of tissues, providing a suitable environment for cells to attach, proliferate, and differentiate. In the current study, we aimed to investigate the effects of decellularized skeletal muscle-derived hydrogel, known as Myogel, on bone marrow-derived MSCs biological behaviors, including proliferation, viability and migration. In this study, MSCs were isolated from tibia and femur of adult Wistar rats. MSCs were cultured in a complete medium (a-MEM containing 15% fetal bovine serum (FBS) and 1% penicillin/streptomycin (Pen/Strep)). The identity of cells was determined by morphology (using inverted microscope) and expression of specific CD markers (using Flowcytometry). Skeletal muscle was decellularized and accuracy of decellularization was evaluated using special staining. Then Myogel was prepared from digested decellularized skeletal muscle. Here, Myogel substrate was used as the control group, gelatin substrate as the positive control, and un-coated plates as the negative control. The effect of Myogel on survival (MTT method), proliferation (drawing the growth curve and calculating the doubling time of the cell population), cell cycle profile (flow cytometry method), and cell migration (scratch method) were investigated. The MTT test showed that the survival of MSCs in Myogel substrate with a concentration of 0.2 mg/ml was higher than the survival of MSCs in gelatin substrate with a concentration of 0.1 mg/ml and the survival of MSCs in gelatin was higher than the survival of control MSCs. Myogel substrate increased the proliferation and migration of cells and decreased the doubling time of MSCs population. Examining the cell cycle profile showed that a high percentage of cells cultured on Myogel were in the G1 and S phase of the cell cycle, indicating an increase in cell division speed by gelatin and, in the next degree, by Myogel. Therefore, Myogel can be used as a suitable substrate to increase the proliferative and migratory potential of MSCs, which are an important factor in tissue engineering.
Development
Fatemeh Ghiyasvand; Somayeh Arabzadeh; mahmood talkhabi
Abstract
Tissue engineering is an emerging field based on the three elements of cells, scaffolds, and bioactive molecules, and can be a useful method for treating muscle injuries. The aim of this study is to investigate the effect of ascorbic acid (AA) on the viability of bone marrow mesenchymal stem cells (BM-MSCs) ...
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Tissue engineering is an emerging field based on the three elements of cells, scaffolds, and bioactive molecules, and can be a useful method for treating muscle injuries. The aim of this study is to investigate the effect of ascorbic acid (AA) on the viability of bone marrow mesenchymal stem cells (BM-MSCs) cultured on skeletal muscle decellularization scaffold. First, BM-MSCs were extracted from rat leg bone marrow and cultured in vitro. The identity of the cells was assessed using flow cytometry. The extracted rat skeletal muscle was decellularized using a 1% SDS solution. The decellularization process was investigated by Masson Trichrome, and Alcian blue and DAPI staining.BM-MSCs were cultured on decellularized scaffolds and treated with 1 mM AA for 2 days. Then, the survival and viability of the cells were evaluated using scanning electron microscope (SEM) and MTT methods, respectively.BM-MSCs had a spindle morphology, and the results of flow cytometry showed the expression of CD44 and CD90 and the lack of expression of CD45 and CD34 in more than 90% of the cells. The staining verified the preservation of collagen and glycosaminoglycans and the absence of DNA in the decellularized tissue. MTT results showed that AA significantly increases the viability of BM-MSCs (P<0.05). Also, the SEM results showed that the cells in the group treated with AA were more proliferated. In general, AA can improve the efficiency of muscle tissue engineering by increasing the viability of BM-MSCs.
Narges Toolabi; Mahmood Talkhabi; Fattane Sam Daliri; Farnoosh Attari; Leila Taghiyar
Abstract
Cancer is the result of the overgrowth of malignant cells that have the ability to spread to other parts of the body. Dichloroacetate (DCA) has been considered as a new drug to control various cancers. The effects of stem cells or their conditioned media (CM) on the treatment or control of some cancers ...
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Cancer is the result of the overgrowth of malignant cells that have the ability to spread to other parts of the body. Dichloroacetate (DCA) has been considered as a new drug to control various cancers. The effects of stem cells or their conditioned media (CM) on the treatment or control of some cancers have also been shown. In this study, human adipose-derived mesenchymal stem cells (h-ADMSCs) and breast cancer cell line 4T1 were first treatedwith different concentrations of DCA and their viability was assessed by MTT assay and 1mM were selected for CM collection. h-ADMSCs with four groups including groups with no FBS media, ± DCA (-FBS /±1mM DCA) and groups with 5% FBS ± DCA (+5%FBS/±1mMDCA) were treated to prepare CM. Then the viability, colony forming potential, cell cycle profile and apoptosis of CM-treated 4T1 cells were investigated. The results showed that CM in the -FBS/+DCA group decreased the viability (P-Value <0.05) and increased the proliferation of 4T1 cells, compared to the -FBS/-DCA group, respectively. Compared to +5%FBS/-DCA group, CM of +5% FBS/+DCA were able to increase viability and proliferation of 4T1 cells. Also, CM of the four studied groups caused changes in the rate of apoptosis and cell cycle profile of 4T1 cells. It seems that, DCA can increase the viability and proliferation of 4T1 breast cancer cells by affecting on thecomposition of mesenchymal stem cells CM.
Mahmood Talkhabi; Hedieh Khanban
Abstract
Mesenchymal stem cells (MSCs) are multipotent stem cells that have potential to differentiate into connective tissue lineages. They also have special properties such as immunomodulation and secretion of growth factors. Histone methyltransferase G9a is one of the factors that control stem cells behaviors ...
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Mesenchymal stem cells (MSCs) are multipotent stem cells that have potential to differentiate into connective tissue lineages. They also have special properties such as immunomodulation and secretion of growth factors. Histone methyltransferase G9a is one of the factors that control stem cells behaviors and features. Hence, it is important to study the role of G9a in MSCs biological behaviors and potentials. MSCs were isolated from rat bone marrow and cultured in vitro. Then, the expression of positive markers (CD90 and CD 73) and negative markers (CD45) were analyzed using flowcytometry. BM-MSCs were treated with different doses of A366, and then were differentiated to osteocyte. Osteogenesis were analyzed using oil red staining and real time-PCR. BM-MSCs were expanded as adherent cells with fibroblastic morphology. More than 85% of cells were positive for CD73 and CD90, and negative for CD45. The treatment of BM-MSCs with A366 reduced osteogenesis as evaluated by oil red staining and gene expression analysis. A366, as an epigenetic regulator decreases the osteogenic potential of BM-MSCs. Use of these regulators for cancer therapy, might influence tissue regeneration and homeostasis.
