scholarly journals Inhibition of G9a Histone Methyltransferase Converts Bone Marrow Mesenchymal Stem Cells to Cardiac Competent Progenitors

2015 ◽  
Vol 2015 ◽  
pp. 1-12 ◽  
Author(s):  
Jinpu Yang ◽  
Keerat Kaur ◽  
Li Lin Ong ◽  
Carol A. Eisenberg ◽  
Leonard M. Eisenberg
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Bone Marrow Cells ◽  
Histone Methyltransferase ◽  
Stem Cell Population ◽  
Molecular Profile ◽  
Sarcomeric Protein ◽  
Marrow Mesenchymal Stem Cells ◽  
Marrow Cells

The G9a histone methyltransferase inhibitor BIX01294 was examined for its ability to expand the cardiac capacity of bone marrow cells. Inhibition of G9a histone methyltransferase by gene specific knockdown or BIX01294 treatment was sufficient to induce expression of precardiac markersMesp1andbrachyuryin bone marrow cells. BIX01294 treatment also allowed bone marrow mesenchymal stem cells (MSCs) to express the cardiac transcription factorsNkx2.5,GATA4, andmyocardinwhen subsequently exposed to the cardiogenic stimulating factor Wnt11. Incubation of BIX01294-treated MSCs with cardiac conditioned media provoked formation of phase bright cells that exhibited a morphology and molecular profile resembling similar cells that normally form from cultured atrial tissue. Subsequent aggregation and differentiation of BIX01294-induced, MSC-derived phase bright cells provoked their cardiomyogenesis. This latter outcome was indicated by their widespread expression of the primary sarcomeric proteins muscleα-actinin and titin. MSC-derived cultures that were not initially treated with BIX01294 exhibited neither a commensurate burst of phase bright cells nor stimulation of sarcomeric protein expression. Collectively, these data indicate that BIX01294 has utility as a pharmacological agent that could enhance the ability of an abundant and accessible stem cell population to regenerate new myocytes for cardiac repair.

scholarly journals Differentiation of Adipose-Derived Mesenchymal Stem Cells into Neuron-Like Cells induced by using β-mercaptoethanol

2020 ◽  
Vol 17 (1(Suppl.)) ◽  
pp. 0235
Author(s):  
Maeda Mohammad ◽  
Ahmed Majeed Al-Shammari ◽  
Rafal H Abdulla ◽  
Aesar Ahmed ◽  
Aseel Khalid
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Light Chain ◽  
Bone Marrow Cells ◽  
Neurofilament Light Chain ◽  
Neurofilament Light ◽  
Study Objective ◽  
Marrow Cells

Background: Adipose derived-mesenchymal stem cells have been used as an alternative to bone marrow cells in this study. Objective: We investigated the in vitro isolation, identification, and differentiation of stem cells into neuron cells, in order to produce neuron cells via cell culture, which would be useful in nerve injury treatment. Method: Mouse adipose mesenchymal stem cells were dissected from the abdominal subcutaneous region. Neural differentiation was induced using β-mercaptoethanol. This study included two different neural stage markers, i.e. nestin and neurofilament light-chain, to detect immature and mature neurons, respectively. Results: The immunocytochemistry results showed that the use of β-mercaptoethanol resulted in the successful production of neuron cells. This was attributable to the increase and significant overexpression of the nestin protein during the early exposure period, which resulted in the expression of the highest levels of nestin. In comparison, the expression level of neurofilament light-chain protein also increased with time but less than nestin. Non-treated mesenchymal stem cells, considered as control showed very low expression for both markers. Conclusion: The results of this study indicate that adipose mesenchymal cells represent a good, easily obtainable source of bone marrow cells used to developing the differentiation process.

Isolation and culture of bone marrow mesenchymal stem cells from human fetus and their biological properties

2008 ◽  
Vol 5 (3) ◽  
pp. 237-244 ◽  
Author(s):  
Zhang Yi-Hua ◽  
Dou Zhong-Ying ◽  
Shen Wen-Zheng ◽  
Yang Chun-Rong ◽  
Gao Zhi-Min
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Calf Serum ◽  
Population Doubling ◽  
Basic Medium ◽  
Long Bones ◽  
Isolated Cells ◽  
Marrow Mesenchymal Stem Cells ◽  
Marrow Cells

AbstractThe population doubling number (70–80 times) of human fetal bone marrow mesenchymal stem cells (BMMSCs) is about two times higher than that (30–40 times) of adult BMMSCs, and their differentiation capacity is superior to that of their adult counterparts. In this study, BMMSCs were isolated from long bones of 2- to 3-month-old human abortuses by rinsing and selectively culturing whole marrow cells. Basic medium and serum concentration were optimized and growth curves plotted, both by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyltetrazolium bromide] reduction assay. Isolated cells were identified by flow cytometry and immunocytochemistry for their antigen markers. The biosafety of isolated cells was evaluated by karyotype analysis and a tumour-forming experiment. The results indicated that lengthwise scissoring of fetal long bones and rinsing of their marrow cells was practical and useful for recovery of BMMSCs from the investigated human abortuses. In this experiment, α-MEM (minimum essential medium, alpha medium)+20% FCS (fetal calf serum) was the best for in vitro culture of BMMSCs. The third-passage BMMSCs expressed Oct4, SSEA3 and SSEA4 besides the surface markers of their adult counterparts. The population doubling time of the BMMSCs of passage 6, 12 and 24 were 34, 36 and 40 h, respectively. Cells in all passages showed a diploid karyotype and formed no tumours in nude mice. The BMMSCs used in this study proved to be biologically safe and ideal seed cells for research on human tissue engineering and regeneration medicine.

Dual effect of WIN-34B on osteogenesis and osteoclastogenesis in cytokine-induced mesenchymal stem cells and bone marrow cells

2016 ◽  
Vol 193 ◽  
pp. 227-236 ◽  
Author(s):  
Byung-Kwan Seo ◽  
Hee-Kyoung Ryu ◽  
Yeon-Cheol Park ◽  
Jeong-Eun Huh ◽  
Yong-Hyeon Baek
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Bone Marrow Cells ◽  
Dual Effect ◽  
Marrow Cells

Mesenchymal Stem Cells of Patients with Chronic Myeloid Leukemia Are Philadelphia Negative.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4683-4683
Author(s):  
Chiara Gentilini ◽  
Kathrin H. Al-Ali ◽  
Annette Reinhardt ◽  
Kristina Bartsch ◽  
Toralf Lange ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Immune Response ◽  
Mesenchymal Stem Cells ◽  
Chronic Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Bone Marrow Cells ◽  
Allogenic Stem Cell Transplantation ◽  
Ph Chromosome ◽  
Marrow Cells

Abstract In the last years, focus of regenerational studies has pointed on mesenchymal stem cells (MSC) and their ability to differentiate into several mesenchymal tissues. MSC have been shown to play a pivotal role in the microenvironment of bone marrow cells and in the modulation of immune response as they can suppress lymphocytic proliferation in vitro. Moreover, some animal studies have suggested they could favor the proliferation of malignant cell clones in solid tumor models. Their role in hematological malignancies, however, remains to be further elucidated and little is known about the influence of MSC in the development and maintenance of the malignant clone in chronic myeloid leukemia (CML). This disease is characterized by the presence of the Philadelphia (Ph) chromosome, a fusion product generated by the reciprocal translocation between chromosomes 9 and 22. Previous reports showed that hepatocytes precursors, found in the liver of CML patients carry the Ph translocation. Our intent was to elucidate whether MSC isolated from patients with CML in different stages of the disease originate from the malignant clone. To this purpose bone marrow aspirates of 11 patients with CML were obtained after informed consent. Five patients were analyzed at diagnosis, two after allogenic stem cell transplantation, three on treatment with the tyrosine kinase inhibitor imatinib and one on treatment with interferon alpha in combination with hydroxyurea. MSC were then generated as previously described. Briefly, cells were isolated by density gradient methods, resuspended in RPMI1640 medium containing 10% fetal bovine serum and plated in culture flasks to adhere. After 4–5 weeks of culture cells were collected and characterized by the expression of several surface markers in a fluorescence activated cell sorter (FACS). The presence of the Ph chromosome was assessed by both fluorescence in situ hybridization (FISH) and polymerase chain reaction (nested PCR). Moreover whole bone marrow was analyzed and results compared with those obtained in the MSC population. MSC showed a typical morphological pattern, growing to confluence after a few weeks of culture and appearing as an adherent, spindle shaped cell layer. In FACS they stained positive for SH2 and SH3 and did not express CD34, CD45 and CD14. MSC were then analyzed by FISH using probes for BCR-ABL. We could not detect the Ph translocation in any of the analyzed patients, though it was present at variuos levels in the remnant bone marrow cells. Results did not change, if expression of BCR-ABL was measured by high sensitivity RT-PCR. Our results showh that MSC of patients with CML are Philadelphia negative irrespective of the stage of disease and the treatment given, suggesting that these cells are not involved in the development of the malignancy. However, their interactions with leukemic cells as well as their role in the immune response against the tumor remains to be further characterized.

In situ delivery of bone marrow cells and mesenchymal stem cells improves cardiovascular function in hypertensive rats submitted to myocardial infarction

2008 ◽  
Vol 15 (3) ◽  
pp. 365-374 ◽  
Author(s):  
Luisa Maria Gomes de Macedo Braga ◽  
Silvia Lacchini ◽  
Beatriz D’Agord Schaan ◽  
Bruno Rodrigues ◽  
Kaleizu Rosa ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Bone Marrow Cells ◽  
Cardiovascular Function ◽  
Hypertensive Rats ◽  
Marrow Cells

Osteogenic Differentiation of Marrow Mesenchymal Stem Cells (MSCs) on Ceramic Total Joints - Significance of Patient's Serum for Proliferation and Differentiation of MSCs -

2006 ◽  
Vol 309-311 ◽  
pp. 1383-1386
Author(s):  
Hajime Ohgushi ◽  
Hiroko Machida ◽  
Akira Oshima ◽  
Noriko Kotobuki ◽  
Motohiro Hirose ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Bone Marrow Cells ◽  
Bone Matrix ◽  
Old Patients ◽  
Proliferation And Differentiation ◽  
Tissue Engineering Approach ◽  
Marrow Mesenchymal Stem Cells

After culture expansion of mesenchymal stem cells (MSCs) from a few milliliter of fresh patient’s bone marrow, we applied the MSCs on alumina ceramic ankle prosthesis and further cultured in an osteogenic medium for 2 weeks. After the culture, the MSCs differentiated into osteoblasts, which fabricated bone matrix on the surface of ceramic prosthesis. The expansion of MSCs followed by osteogenic differentiation was done using the commercially available medium with some chemicals and patient’s own serum. The MSCs well proliferated and differentiated into osteoblasts, even the MSCs were from old aged (more than 70 years old) patients. The tissue engineered ceramic prostheses were implanted into osteoarthritic patients. Typical X-ray findings showed that radiodense areas began to appear around the cell-seeded areas on the prosthesis about 2 to 3 months after the operation. These findings confirmed the importance of tissue engineering approach for early bone fixation and the approach can be done using small number of bone marrow cells and patient’s own serum without adding animal-derived products.

Human mesenchymal stem cells xenografted directly to rat liver are differentiated into human hepatocytes without fusion

Blood ◽  
2005 ◽  
Vol 106 (2) ◽  
pp. 756-763 ◽  
Author(s):  
Yasushi Sato ◽  
Hironobu Araki ◽  
Junji Kato ◽  
Kiminori Nakamura ◽  
Yutaka Kawano ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Rat Liver ◽  
Bone Marrow Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Hepatic Differentiation ◽  
Cd34 Cells ◽  
Marrow Cells

Abstract Hepatic transdifferentiation of bone marrow cells has been previously demonstrated by intravenous administration of donor cells, which may recirculate to the liver after undergoing proliferation and differentiation in the recipient's bone marrow. In the present study, to elucidate which cellular components of human bone marrow more potently differentiate into hepatocytes, we fractionated human bone marrow cells into mesenchymal stem cells (MSCs), CD34+ cells, and non-MSCs/CD34- cells and examined them by directly xenografting to allylalcohol (AA)-treated rat liver. Hepatocyte-like cells, as revealed by positive immunostaining for human-specific alpha-fetoprotein (AFP), albumin (Alb), cytokeratin 19 (CK19), cytokeratin 18 (CK18), and asialoglycoprotein receptor (AGPR), and by reverse transcription-polymerase chain reaction (RT-PCR) for expression of AFP and Alb mRNA, were observed only in recipient livers with MSC fractions. Cell fusion was not likely involved since both human and rat chromosomes were independently identified by fluorescence in situ hybridization (FISH). The differentiation appeared to follow the process of hepatic ontogeny, reprogramming of gene expression in the genome of MSCs, as evidenced by expression of the AFP gene at an early stage and the albumin gene at a later stage. In conclusion, we have demonstrated that MSCs are the most potent component in hepatic differentiation, as revealed by directly xenografting into rat livers. (Blood. 2005;106:756-763)

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