Bone Morphogenetic Protein Signaling Is Involved in Human Mesenchymal Stem Cell Survival in Serum-Free Medium

Solmesky, Leonardo J.; Abekasis, Michal; Bulvik, Shlomo; Weil, Miguel

doi:10.1089/scd.2009.0020

Cited by 16 publications

(20 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Moreover, as mentioned above we also characterized hMSC by their biological property of prolonged survival under serum free conditions [7,21]. To this end ALS hMSC and non-ALS hMSC were cultured at low and high densities of 20 and 60 cells per square millimeter in 24 well dishes, respectively in DMEM with or without 10% serum.…”

Section: Resultsmentioning

confidence: 99%

“…To this end ALS hMSC and non-ALS hMSC were cultured at low and high densities of 20 and 60 cells per square millimeter in 24 well dishes, respectively in DMEM with or without 10% serum. Cell viability in each well was determined at the end of the incubation period of 1, 2 and 3 days using a vital fluorescent dye mix containing Calcein-AM (stains living cells), Propidium iodide (PI) (stains dead cells) and Hoechst 33342 (stains all nuclei) as described previously [7]. Figure 1 shows representative results from these characterization experiments of hMSC.…”

Section: Resultsmentioning

confidence: 99%

“…hMSC were isolated and cultured as described before [7]. hMSC at passages 3-7 were used in this study.…”

Section: Isolation and Culture Of Hmscmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…Over 100 distinct mu- [6]. Moreover, previous study in our laboratory reveals that these stem cells bearthe rare biological capacity to survive for prolonged time under serum free culture conditions due to the production of their own survival factors, like bone morphogenetic protein [7].RNA editing is a post-transcriptional mechanism for expanding the proteomic repertoire. Adenosineto-inosine (A to I) RNA editing by enzymes named ADARs (adenosine deaminases acting on RNAs) is essential for normal life and development of both invertebrates and vertebrates [8][9][10].…”

mentioning

confidence: 99%

See 4 more Smart Citations

Two Potential Biomarkers Identified in Mesenchymal Stem Cells and Leukocytes of Patients with Sporadic Amyotrophic lateral Sclerosis

et al. 2012

Self Cite

View full text Add to dashboard Cite

Abstract. Amyotrophic lateral sclerosis (ALS) is a fatal, neurodegenerative disorder caused by degeneration of motor neurons.The cause for most cases of ALS is multi-factorial,this enhances the need to characterize and isolate specific biomarkers found in biological samples from ALS patients. To this end we use human mesenchymal stem cells (hMSC) derived from the bone marrow of six ALS patients (ALS hMSC) and identified two genes, Cytoplasmic FMR Interacting Protein 2 (CyFIP2) and Retinoblastoma (Rb) Binding Protein 9 (RbBP9) with a significant decrease in post transcriptional A to I RNA editing compared to hMSC of healthy individuals. At the transcriptional level we show abnormal expression of these two genes in ALS hMSC by quantitative real time-PCR (qRT-PCR) and Western blot suggesting a problem in the regulation of these genes in ALS. To strengthen this view we tested by qRT-PCR the expression of these genes in peripheral blood leukocytes (PBL) isolated from blood samples of 17 ALS patients and found that CyFIP2 and RbBP9 levels of expression were significantly different compared to the levels of expression of these two genes in 19 normal PBL samples. Altogether we found two novel ALS potential biomarkers in non-neural tissues from ALS patients that may have direct diagnostic and therapeutic implications to the disease.Keywords: ALS biomarkers, CyFIP2, RbBP9, human mesenchymal stem cells, leukocytes BackgroundALS is a fatal and incurable neurodegenerative disorder characterized by progressive loss of motor neurons in the spinal cord, brainstem and motor cortex, resulting in generalized weakness and muscle atrophy. ALS is the most common motor neuron disorder, with a prevalence of 6 per 100,000 at any given time [1]. Approximately 90% of the cases are sporadic, and the remaining 10% are familial [2]. Over 100 distinct mu- [6]. Moreover, previous study in our laboratory reveals that these stem cells bearthe rare biological capacity to survive for prolonged time under serum free culture conditions due to the production of their own survival factors, like bone morphogenetic protein [7].RNA editing is a post-transcriptional mechanism for expanding the proteomic repertoire. Adenosineto-inosine (A to I) RNA editing by enzymes named ADARs (adenosine deaminases acting on RNAs) is essential for normal life and development of both invertebrates and vertebrates [8][9][10]. When functioning improperly, this essential process, can lead to pathological conditions ranging from epilepsy to malignant gliomas [11][12][13]. So far, only abnormal RNA editing at the Q/R (glutamine/arginine) site of the GluR-2 gene of the AMPA (Amino-3-hydroxy-5-methyl-4-isoxazolepropionate) receptor has been identified to be implicated in ALS [14][15][16][17]. It is unknown whether the RNA editing deficiency in the GluR-2 gene is part of a more global RNA problem related with ADARs activity in ALS [18].Here we show that ALS hMSC have abnormal pattern of RNA editing in two novel genes, CyFIP2 and RbBP9, which also expressed significantly differen...

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

“…hMSC were isolated and cultured as described before [7]. hMSC at passages 3-7 were used in this study.…”

Section: Isolation and Culture Of Hmscmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Two Potential Biomarkers Identified in Mesenchymal Stem Cells and Leukocytes of Patients with Sporadic Amyotrophic lateral Sclerosis

et al. 2012

Self Cite

View full text Add to dashboard Cite

show abstract

BMP2 induces osteoblast apoptosis in a maturation state and noggin‐dependent manner

Hyzy

Olivares-Navarrete

Schwartz

et al. 2012

J of Cellular Biochemistry

View full text Add to dashboard Cite

Large doses of bone morphogenetic protein 2 (BMP2) are used clinically to induce bone formation in challenging bone defects. However, complications after treatment include swelling, ectopic bone formation, and adjacent bone resorption. While BMP2 can be effective, it is important to characterize the mechanism of the deleterious effects to optimize its use. The aim of this study was to determine the effect of BMP2 on apoptosis in osteoblast lineage cells and to determine the role of the BMP inhibitor Noggin in this process. Human mesenchymal stem cells (MSCs), immature osteoblast-like MG63 cells, and mature normal human osteoblasts (NHOst) were treated with BMP2. A model system of increased endogenous BMP signaling was created by silencing Noggin (shNOG-MG63). Finally, the BMP pathway regulating apoptosis in NHOst was examined using BMP signaling inhibitors (5Z-7-oxozeaenol, dorsomorphin, H-8). Apoptosis was characterized by caspase-3, BAX/BCL2, p53, and DNA fragmentation. BMP2 induced apoptosis in a cell-type dependent manner. While the effect was minor in MSCs, MG63 cells had modest increases and NHOst cells had robust increases apoptosis after BMP2 treatment. Apoptosis was significantly higher in shNOG-MG63 than MG63 cells. 5Z-7-oxozeaenol and dorsomorphin eliminated the BMP2-induced increase in DNA fragmentation in NHOst, suggesting roles for TAB/TAK1 and Smad signaling. These results indicate that the apoptotic effect of BMP2 is dependent on cell maturation state, inducing apoptosis in committed osteoblasts through Smad and TAB/TAK1 signaling, and is regulated by Noggin. Dose and delivery must be optimized in therapeutic applications of BMP2 to minimize complications.

show abstract

Adenosine Triphosphate Prevents Serum Deprivation-Induced Apoptosis in Human Mesenchymal Stem Cells via Activation of the MAPK Signaling Pathways

et al. 2014

View full text Add to dashboard Cite

Human mesenchymal stem cells (hMSC) are multipotent cells derived from various sources including adipose and placental tissues as well as bone marrow. Owing to their regenerative and immunomodulatory properties, their use as a potential therapeutic tool is being extensively tested. However, one of the major hurdles in using cell-based therapy is the use of fetal bovine serum that can trigger immune responses, viral and prion diseases. The development of a culture medium devoid of serum while preserving cell viability is therefore a major challenge. In this study, we demonstrated that adenosine triphosphate (ATP) restrained serum deprivation-induced cell death in hMSC by preventing caspases 3/7 activation and modulating ERK1/2 and p38 MAPK signaling pathways. We also showed that serum deprivation conditions triggered dephosphorylation of the proapoptotic protein Bad leading to cell death. Adjunction of ATP restored the phosphorylation state of Bad. Furthermore, ATP significantly modulated the expression of proapoptopic and antiapoptotic genes, in favor of an antiapoptotic profile expression. Finally, we established that hMSC released a high amount of ATP in the extracellular medium when cultured in a serum-free medium. Collectively, our results demonstrate that ATP favors hMSC viability in serum deprivation conditions. Moreover, they shed light on the cardinal role of the MAPK pathways, ERK1/2 and p38 MAPK, in promoting hMSC survival. STEM CELLS 2015;33:211-218

show abstract

Bone Morphogenetic Protein Signaling Is Involved in Human Mesenchymal Stem Cell Survival in Serum-Free Medium

Cited by 16 publications

References 21 publications

Two Potential Biomarkers Identified in Mesenchymal Stem Cells and Leukocytes of Patients with Sporadic Amyotrophic lateral Sclerosis

Two Potential Biomarkers Identified in Mesenchymal Stem Cells and Leukocytes of Patients with Sporadic Amyotrophic lateral Sclerosis

BMP2 induces osteoblast apoptosis in a maturation state and noggin‐dependent manner

Adenosine Triphosphate Prevents Serum Deprivation-Induced Apoptosis in Human Mesenchymal Stem Cells via Activation of the MAPK Signaling Pathways

Contact Info

Product

Resources

About