Characterization of mesenchymal stem cells from rat bone marrow: ultrastructural properties, differentiation potential and immunophenotypic markers

2009 ◽  
Vol 132 (5) ◽  
pp. 533-546 ◽  
Author(s):  
Erdal Karaoz ◽  
Ayça Aksoy ◽  
Selda Ayhan ◽  
Ayla Eker Sarıboyacı ◽  
Figen Kaymaz ◽  
...  
2021 ◽  
Vol 11 (5) ◽  
pp. 957-962
Author(s):  
Ainiwaerjiang Damaola ◽  
Maerdan Aierken ◽  
Mieralimu Muertizha ◽  
Abudouaini Abudoureheman ◽  
Haishan Lin ◽  
...  

We aimed to explore the effects of rat bone marrow mesenchymal stem cells (BMSCs) on osteogenic differentiation via analyzing miR-3148 expression in patients with osteoporosis. Realtime quantitative PCR was conducted for assessing microRNA-3148 expression. BMSCs from SD rats were transfected with microRNA-3148 mimics and microRNA-3148 inhibitor via liposomal trans-fection method utilizing Lipo2000, followed by analysis of microRNA-3148 level. After 10-days of osteogenic differentiation induction, alkaline phosphatase (ALP) staining and alizarin red (ARS) staining were done to investigate the osteogenic differentiation potential. Simultaneously, qRT-PCR measured the expression of osteogenesis marker genes (BMP and Runx2) in each group. qRT-PCR analysis revealed a high expression of miR-3148 in the bone tissue and the serum samples from patients with osteoporosis in comparison with healthy individuals. In addition, miRNA-3148 mimics could retard the osteogenic differentiation of BMSCs, while microRNA-3148 inhibitor could prompt the procedure. MicroRNA-3148 was highly expressed in the skeletal tissues and the serum samples from patients with osteoporosis and it could restrain the differentiation of BMSCs into osteoblasts, suggesting that it might be a novel therapeutic target for treating osteoporosis.


2011 ◽  
Vol 34 (2) ◽  
pp. 281-290 ◽  
Author(s):  
Samy Hosny Hammed ◽  
Amany Mohamed El Shawarby ◽  
Mohamed Abd Elrahman Ahmed ◽  
Mohamed Kamel Abo Golayel ◽  
Asmaa Abd Elmonem Mohamed

2017 ◽  
Vol 4 (S) ◽  
pp. 25
Author(s):  
Karuppiah Thilakavathy

Preclinical studies on mesenchymal stem cells (MSC) have allowed the cells to be considered as a promising candidate for cellular therapy. The mouse is the most widely used species for studying the characteristics of MSC. In recent years, conflicting data were reported regarding growth kinetics, surface marker profile, differentiation capacity, genetic instability or malignant transformation and so forth, that may be a result of a range of factors. One of the factors probably is the culture medium formulation.  Here we have made a comparative characterization of bone marrow-derived mesenchymal stem cells (mBM-MSC), under the same experimental conditions, cultured using two common supplements, fetal bovine serum (FBS) and MesenCultTM Stimulatory Supplement (MSS). mBM-MSC isolated from the tibias of C57BL/6 mice were cultured and expanded in Dulbecco’s Modified Eagle’s Medium supplemented with either 15% FBS or 15% MSS.  Clonogenic potential, population doubling time, immunophenotyping, differentiation immunosuppression potentials and chromosome analysis of early and late passage of mBM-MSC were assessed.      The findings showed that the immunophenotype and differentiation potential of mBM-MSC were similar when cultured using these supplements irrespective of passages.  Variations were seen in clonogenic, growth, proliferation rate and immunosuppression potential of the mBM-MSC.  This study also revealed that prolonged culture will disrupt their genetic stability regardless of the supplements used.  The genetically mutated mBM-MSC were also found to maintain their stemness characteristics and immunosuppression potential.       In conclusion, culture medium formulation causes variations in the cultured MSC and may influence downstream investigation findings.


2014 ◽  
Vol 7 (2) ◽  
pp. 135-142 ◽  
Author(s):  
Ahmed Lotfy ◽  
Mohamed Salama ◽  
Faten Zahran ◽  
Elena Jones ◽  
Ahmed Badawy ◽  
...  

2013 ◽  
Vol 65 (4) ◽  
pp. 1307-1315
Author(s):  
Alexandra Ivan ◽  
V. Ordodi ◽  
Ada Cean ◽  
Daniela Ilie ◽  
Carmen Panaitescu ◽  
...  

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4260-4260
Author(s):  
Maria G. Roubelakis ◽  
Kalliopi I. Pappa ◽  
Vassiliki Bitsika ◽  
Dimitra Zagoura ◽  
Antonia Vlahou ◽  
...  

Abstract Human mesenchymal stem cells (hMSCs) constitute a population of multipotent cells, easily expanded in culture and able to give rise to many lineages. These characteristics make MSCs a very attractive tool for developing new strategies for clinical applications based on cell therapy. So far, the most common source of MSCs has been the bone marrow (BM). However, identification and characterization of alternative sources of MSCs is of great importance. One such alternative source is the amniotic fluid (AF), which can be collected during scheduled amniocentesis without any ethical concerns. To this end, in the present study, we introduced an improved protocol for isolating and clonally expanding fetal MSCs from second trimester amniotic fluid (AF) and we further characterized these cells based on their phenotype, pluripotency, differentiation potential and proteomic profile. The AF samples were obtained during routine amniocentesis and AF-MSCs were enriched by a modified culture protocol. The isolated MSCs expanded rapidly and exhibited differentiation potential into adipocytes and osteoblasts. More importantly, we showed that these cells can differentiate in vitro not only into cell types derived from mesoderm (adipocytes and osteoblasts) and ectoderm (neural cells) but also more interestingly into endoderm (hepatocytes) derived cells. Moreover, we documented that AF-MSCs express Oct-4 transcription factor, a marker of pluripotency, and we studied for the first time its expression over different passages by real time PCR and documented that it remained constant for at least 17 doublings. An extensive characterization of the phenotypic features of AF-MSCs by using a wide range of surface markers and flow cytometry, indicated that they are positive for all the mesenchymal stem cell markers such as CD90, CD105, CD73 and CD166 and generally exhibit a similar expression pattern to the BM-MSCs. To characterize these cells in more detail, we established the first proteomic database for human AF-MSCs. Using 2D-gel electrophoresis and matrix-assisted laser desorption ionisation-time of flight-mass (MALDI-TOF) spectrometry approach, we have generated for the first time the protein map of AF MSCs, by identifying 260 proteins and directly compared this protein profile with that of MSCs derived from BM. We further performed a similar analysis for BM-MSCs, identifying 170 different proteins and generating a reference map for these cells. The comparison of the proteomic pattern from both sources was similar. In general, 140 proteins were identified in AF-MSCs related to cell growth/maintenance, metabolism/energy pathways, protein metabolism, apoptosis, signal transduction and communication as well as transcription and transport, that are not present in BM-MSCs. The approach we initiated, is expected to facilitate systematic functional studies for these multipotent cells. One such approach could be the implementation of the proteomic analysis, during differentiation of AF-MSCs to cells derived from all three germ layers as shown in our study. Data derived from these approaches are expected to clarify the therapeutic potential of the MSCs.


2018 ◽  
Vol 18 ◽  
Author(s):  
Chaitra Venugopal ◽  
Christopher Shamir ◽  
Sivapriya Senthilkumar ◽  
Janitri Venkatachala Babu ◽  
Peedikayil Kurien Sonu ◽  
...  

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