Long bone mesenchymal stem cells (Lb-MSCs): clinically reliable cells for osteo-diseases

Toosi, Shirin; Naderi‐Meshkin, Hojjat; Kalalinia, Fatemeh; Pievandi, Mohammad Taghi; Hosseinkhani, Hossein; Bahrami, Ahmad Reza; Heirani‐Tabasi, Asieh; Mirahmadi, Mahdi; Behravan, Javad

doi:10.1007/s10561-017-9652-3

Cited by 24 publications

(14 citation statements)

References 46 publications

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“…The RIA device is currently used to fix long bones with intramedullary nails. The ICBM that is taken in this process is wasted and several studies have shown that aspirates from the reaming system have large amounts of MSCs (Toosi et al, 2017;van der Bel & Blokhuis, 2014;Wenisch et al, 2005 (Porter et al, 2009). Cox et al (2011) (Bourin et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Adipocyte lineage differentiation potential of MSCs isolated from reaming material

Toosi

Esmaeilzadeh

Naderi‐Meshkin

et al. 2019

Journal Cellular Physiology

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Mesenchymal stem cells (MSCs) obtained from various sources have been used for different therapeutic applications including tissue regeneration. Reamer/irrigator/ aspirator (RIA) has been increasingly used in recent years for the derivation of MSCs.Here in this investigation we have comparatively analyzed MSCs obtained from iliac crest bone marrow (ICBM) and RIA for their morphology, cluster determinant (CD) markers, and adipogenic differentiation capacity. MSCs were isolated, cultured, and purified from both sources and then flow cytometric studies were performed to study their characteristics. The differentiation potential of RIA and ICBM was examined by an Oil Red O staining protocol. Moreover, the tissue-specific markers related to adipogenesis were analyzed by real-time polymerase chain reaction (RT-PCR). The cells were cultured in the relevant induction medium and then adipogenic lineage differentiation was tested and confirmed for all MSC preparations. Additionally, analysis by flow cytometer was indicative of RIA derived MSCs (RIA-MSCs) having a more homogenous population than ICBM derived MSCs. The RIA-MSCs differentiation toward adipogenic lineage was more efficient compared with ICBM-MSCs. Direct comparative analysis of RIA to ICBM-MSCs indicated that the RIA-MSCs had a higher potential toward adipocyte lineage differentiation compared with ICBM-MSCs.

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Section: Discussionmentioning

confidence: 99%

Adipocyte lineage differentiation potential of MSCs isolated from reaming material

Toosi

Esmaeilzadeh

Naderi‐Meshkin

et al. 2019

Journal Cellular Physiology

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“…The pellet was centrifuged again at 400 g for 6 minutes, resuspended and cultivated in low glucose Dulbecco's modified Eagle’s medium (DMEM) (Biowest, Nuaillé, France) composed of 2 ng/ml basic fibroblast growth factor (Royan Institute, Tehran, Iran), 10% fetal bovine serum (FBS) (Invitrogen), 100 mg/ml streptomycin and 100 U/ml penicillin (Invitrogen) . 33,34 …”

Section: Methodsmentioning

confidence: 99%

CRISPR/Cas9 mediated GFP‐human dentin matrix protein 1 (DMP1) promoter knock‐in at the ROSA26 locus in mesenchymal stem cell for monitoring osteoblast differentiation

Shahabipour

Oskuee

Shokrgozar

et al. 2020

The Journal of Gene Medicine

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Background: Dentin matrix protein 1 (DMP1) is highly expressed in mineralized tooth and bone, playing a critical role in mineralization and phosphate metabolism. One important role for the expression of DMP1 in the nucleus of preosteoblasts is the up-regulation of osteoblast-specific genes such as osteocalcin and alkaline phosphatase 1. The present study aimed to investigate the potential application of human DMP1 promoter as an indicator marker of osteoblastic differentiation. Methods: In the present study, we developed DMP1 promoter-DsRed-GFP knock-in mesenchymal stem cell (MSCs) via the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system that enabled automatic detection of osteoblast differentiation. With the application of a homology-directed knock-in strategy, a 2-kb fragment of DMP1 promoter, which was inserted upstream of the GFP and DsRed reporter cassette, was integrated into the human ROSA locus to generate double fluorescent cells. We further differentiated MSCs under osteogenic media to monitor the fate of MSCs. First, cells were transfected using CRISPR/Cas9 plasmids, which culminated in MSCs with a green fluorescence intensity, then GFP-positive cells were selected using puromycin. Second, the GFP-positive MSCs were differentiated toward osteoblasts, which demonstrated an increased red fluorescence intensity. The osteoblast differentiation of MSCs was also verified by performing alkaline phosphatase and Alizarin Red assays. Results: We have exploited the DMP1 promoter as a predictive marker of MSC differentiation toward osteoblasts. Using the CRISPR/Cas9 technology, we have identified a distinctive change in the fluorescence intensities of GFP knock-in (green) and osteoblast differentiated MSCs 2 .

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“…In following of osteogenic cultures were stained with BCIP/NBT tablet (Becton Dickinson, Bioscience) and Alizarin Red S (Sigma-Aldrich). Also, the fat vacuoles in the adipogenic cultures were stained with Oil Red O (Sigma-Aldrich) (Toosi et al 2017). Additionally, Ad-MSCs were analysed by flow cytometry for their Cluster Differentiation (CD) markers.…”

Section: Cell Culture and Characterizationmentioning

confidence: 99%

Comparison the effects of hypoxia-mimicking agents on migration-related signaling pathways in mesenchymal stem cells

et al. 2020

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Adipose-derived mesenchymal stem cells (Ad-MSCs) have been designated as the promising agents for clinical applications for easy accessibility, multi-linage differentiation and immunomodulation capacity. Despite this, optimal cell delivery conditions have remained as a clinical challenge and improvement of stem cell homing to the target organs is being considered as a major strategy in cell therapy systemic injection. It has been shown that homing of mesenchymal stem cells are increased when treated with physical or chemical hypoxia-mimicking factors, however, efficiency of different agents remained to be determined. In this study, hypoxia-mimicking agents, including valproic acid (VPA), cobalt chloride (CoCl 2) and deferoxamine (DFX) were examined to determine whether they are able to activate signaling molecules involved in migration of Ad-MSCs in vitro. We report that Ad-MSCs treated by DFX resulted in a significantly enhanced mRNA expression of MAPK4 (associated with MAPK signaling pathway), INPP4B (associated with Inositol polyphosphate pathway), VEGF-A and VEGF-C (associated with cytokinecytokine receptor pathways), IL-8 and its receptor, CXCR2 (associated with IL-8 signaling pathway). While the cells treated with VPA did not show such effects and CoCl 2 only upregulated VEGF-A and VEGF-C gene expression. Furthermore, results of wound-healing assays showed migration capacity of Ad-MSCs treated with DFX significantly increased 8 and 24 h of the treatment. This study provides credible evidence around DFX, which might be an effective drug for pharmacological preconditioning of Ad-MSCs to boost their homing capacity and regeneration of damaged tissues though, activation of the migration-related signaling pathways.

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Long bone mesenchymal stem cells (Lb-MSCs): clinically reliable cells for osteo-diseases

Cited by 24 publications

References 46 publications

Adipocyte lineage differentiation potential of MSCs isolated from reaming material

Adipocyte lineage differentiation potential of MSCs isolated from reaming material

CRISPR/Cas9 mediated GFP‐human dentin matrix protein 1 (DMP1) promoter knock‐in at the ROSA26 locus in mesenchymal stem cell for monitoring osteoblast differentiation

Comparison the effects of hypoxia-mimicking agents on migration-related signaling pathways in mesenchymal stem cells

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