Farid Ebnerasuly1,2, Zahra Hajebrahimi3, Seyed Mehdi Tabaie4, Mojtaba Darbouy1,2. 1. Department of Biology, Fars Science and Research Branch , Islamic Azad University, Marvdasht, Iran. 2. Department of Biology, Marvdasht Branch, Islamic Azad University, Marvdasht, Iran. 3. Aerospace Research Institute, Ministry of Science Research and Technology, Tehran, Iran. 4. Medical Laser Research Center, Iranian Academic Center for Education, Culture and Research (ACECR), Tehran, Iran.
Abstract
Background: Mesenchymal stem cells (MSCs) are multipotent cells able to differentiating into a variety of mesenchymal tissues including osteoblasts, adipocytes and several other tissues. Objectives: Differentiation of MSCs into fibroblast cells in vitro is an attractive strategy to achieve fibroblast cell and use them for purposes such as regeneration medicine. The goal of this study was investigate the simulated microgravity effect on differentiation of Adipose Derived Stem Cells (ADSCs) to fibroblasts. Materials and Methods: To fibroblast differentiation 100 ng.mL-1 of connective tissue growth factor (CTGF), and for simulation microgravity, 2D clinostat was used. After isolation the human ADSCs from adipose, cells were passaged, and at passages 3 they were used for characterization and subsequent steps. After 7 days of CTGF and simulated microgravity treatment, proliferation, and differentiation were analyzed collectively by MTT assay, quantitative PCR analyses, and Immunocytochemistry staining. Results: MTT assay revealed that CTGF stimulate the proliferation but simulated microgravity didn't have statistically significant effect on cell proliferation. In RNA level the expression of these genes are investigated: collagen type I (COLI), elastin (ELA), collagen type III (ColIII), Matrix Metalloproteinases I(MMP1), Fibronectin 1 (FN1), CD44, Fibroblast Specific protein (FSP-1), Integrin Subunit Beta 1 (ITGB1), Vimentin (VIM) and Fibrillin (FBN). We found that expression of ELN, FN1, FSP1, COL1A1, ITGB1, MMP1 and COL3A1 in both condition, and VIM and FBN1 just in differentiation medium in normal gravity increased. In protein level the expression of COL III and ELN in simulated microgravity increased. Conclusions: These findings collectively demonstrate that the simulated microgravity condition alters the marker fibroblast gene expression in fibroblast differentiation process.
Background: Mesenchymal stem cells (MSCs) are multipotent cells able to differentiating into a variety of mesenchymal tissues including osteoblasts, adipocytes and several other tissues. Objectives: Differentiation of MSCs into fibroblast cells in vitro is an attractive strategy to achieve fibroblast cell and use them for purposes such as regeneration medicine. The goal of this study was investigate the simulated microgravity effect on differentiation of Adipose Derived Stem Cells (ADSCs) to fibroblasts. Materials and Methods: To fibroblast differentiation 100 ng.mL-1 of connective tissue growth factor (CTGF), and for simulation microgravity, 2D clinostat was used. After isolation the human ADSCs from adipose, cells were passaged, and at passages 3 they were used for characterization and subsequent steps. After 7 days of CTGF and simulated microgravity treatment, proliferation, and differentiation were analyzed collectively by MTT assay, quantitative PCR analyses, and Immunocytochemistry staining. Results:MTT assay revealed that CTGF stimulate the proliferation but simulated microgravity didn't have statistically significant effect on cell proliferation. In RNA level the expression of these genes are investigated: collagen type I (COLI), elastin (ELA), collagen type III (ColIII), Matrix Metalloproteinases I(MMP1), Fibronectin 1 (FN1), CD44, Fibroblast Specific protein (FSP-1), Integrin Subunit Beta 1 (ITGB1), Vimentin (VIM) and Fibrillin (FBN). We found that expression of ELN, FN1, FSP1, COL1A1, ITGB1, MMP1 and COL3A1 in both condition, and VIM and FBN1 just in differentiation medium in normal gravity increased. In protein level the expression of COL III and ELN in simulated microgravity increased. Conclusions: These findings collectively demonstrate that the simulated microgravity condition alters the marker fibroblast gene expression in fibroblast differentiation process.
Entities:
Keywords:
Adipose Derived Stem Cells (ADSCs); Differentiation; Extracellular Matrix (ECM); Fibroblast; Real Time PCR; Simulated Microgravity
Authors: Gregory H Altman; Rebecca L Horan; Ivan Martin; Jian Farhadi; Peter R H Stark; Vladimir Volloch; John C Richmond; Gordana Vunjak-Novakovic; David L Kaplan Journal: FASEB J Date: 2001-12-28 Impact factor: 5.191
Authors: Wolfgang Wagner; Frederik Wein; Anja Seckinger; Maria Frankhauser; Ute Wirkner; Ulf Krause; Jonathon Blake; Christian Schwager; Volker Eckstein; Wilhelm Ansorge; Anthony D Ho Journal: Exp Hematol Date: 2005-11 Impact factor: 3.084
Authors: Xiaoqing Guo; Audrey E K Hutcheon; Suzanna A Melotti; James D Zieske; Vickery Trinkaus-Randall; Jeffrey W Ruberti Journal: Invest Ophthalmol Vis Sci Date: 2007-09 Impact factor: 4.799
Authors: Kimberly G Riley; Raymond C Pasek; Matthew F Maulis; Jennifer Peek; Fabrizio Thorel; David R Brigstock; Pedro L Herrera; Maureen Gannon Journal: Diabetes Date: 2014-11-12 Impact factor: 9.461
Authors: Daniela Grimm; Markus Wehland; Thomas J Corydon; Peter Richter; Binod Prasad; Johann Bauer; Marcel Egli; Sascha Kopp; Michael Lebert; Marcus Krüger Journal: Stem Cells Transl Med Date: 2020-04-30 Impact factor: 6.940
Authors: Zheng-Yang Chen; Song Guo; Bin-Bin Li; Nan Jiang; Ao Li; Hong-Feng Yan; He-Ming Yang; Jin-Lian Zhou; Cheng-Lin Li; Yan Cui Journal: Biomed Res Int Date: 2019-04-03 Impact factor: 3.411