Behrouz Zandieh Doulabi |
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PhD
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Academisch Centrum Tandheelkunde Amsterdam
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Field of Oral Cell Biology
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Skills (5)
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78 Questions1404 Followers
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0 Questions10 Followers
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109 Questions8001 Followers
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123 Questions15370 Followers
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241 Questions25854 Followers
Research experience
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Jan 2009–
Dec 2012Research: VU University Amsterdam
VU University Amsterdam · Oral Cell Biology and Functional AnatomyNetherlands · Amsterdam -
Jan 2007–
Dec 2011Research: VU medisch centrum
VU medisch centrum · Department of Orthopedic SurgeryNetherlands · Amsterdam -
Jan 2006–
Dec 2009Research: Academisch Centrum Tandheelkunde Amsterdam
Academisch Centrum Tandheelkunde AmsterdamNetherlands · Amsterdam -
Jan 2002–
Dec 2012Research: Universiteit van Amsterdam
Universiteit van Amsterdam · Faculty of Medicine AMCNetherlands · Amsterdam -
Jan 2001–
Dec 2004Research: Academisch Medisch Centrum Universiteit van Amsterdam
Academisch Medisch Centrum Universiteit van Amsterdam · Department of EndocrinologyNetherlands · Amsterdam -
Jan 1998
Research: Agricultural University Wageningen
Netherlands · Wageningen
Education
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Jan 1999–
Jan 2003Universiteit van Amsterdam
PhDNetherlands · Amsterdam -
Sep 1994–
Jul 1996Universiteit van Amsterdam
MSCNetherlands · Amsterdam -
Sep 1987–
Jun 1991Hogeschool van Amsterdam
BSCAmsterdam
Other
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LanguagesDutch, English, Persian
Questions and Answers (24) View all
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Answer added in Cell Culture151 What are the best kits for RNA isolation?By Danielle Smith · University of Cape TownBehrouz Zandieh Doulabi · Academisch Centrum Tandheelkunde AmsterdamDear Danielle, Hi. Regarding to question I am afraid to say that the best general kit for RNA isolation does not exist. Choice of kits is first depend... [more]Dear Danielle, Hi. Regarding to question I am afraid to say that the best general kit for RNA isolation does not exist. Choice of kits is first depending on downstream assays (Northern blot, microarrays, Real time PCR from total RNA, mRNA or microRNA). In general there are 3 approaches 1. Phenol extraction based methods (such as Trizol or Tri pure) which are cheap and are ok for microarray analysis and isolating total RNA and microRNAs . However, disadvantage remains contamination with proteins or genomic DNA and or phenol and other solvent residues inhibiting cDNA or DNAsynthesis related enzymes such as Taq polymerases. 2. Affinity column based methods such as Qiagen depending type of cell or tissues, Highpure from Roche or nucleospin (if you have not much cells; 4000 cells and more). The quality of RNA is fair and RNA is clean however, it cannot always used for some applications e.g efficient microRNAs isolation and yield is lower than other two methods. 3. Magnetic beads based methods such as Roche Magnapure which needs robot to isolate RNA or dynal from Life Sciences. This method ensures best quality and yield for mRNA or total RNA isolations. However, the price is the most disadvantage of using magnetic beads. I hope you can use this information for choice of best working method for your specific research questionFollowing
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Answer added in ELISA8 Will protein isolated with Nucleospin RNA/Protein kit work when used in ELISA?By Mohammed Aliyu-Paiko · Putra University, MalaysiaBehrouz Zandieh Doulabi · Academisch Centrum Tandheelkunde AmsterdamDear Mohammed, Nucleospin kit is actually designed for Western blot analysis and denaturate and reduces proteins completely. Even after dialysing and... [more]Dear Mohammed, Nucleospin kit is actually designed for Western blot analysis and denaturate and reduces proteins completely. Even after dialysing and or reprecipation is not easy to use the protein for ElISA. I would suggest to try Western blots with antibodies recognizing the denatured and reduced protein. Good LuckFollowing
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Answer added in PCR13 How can we have an amplification efficiency greater than 100%?By Virginie Mournetas · University of LiverpoolBehrouz Zandieh Doulabi · Academisch Centrum Tandheelkunde AmsterdamDear Virginie I suspect that you have constant forming and dissociating primer dimmers and because you have more primer dimer molecules then your spec... [more]Dear Virginie I suspect that you have constant forming and dissociating primer dimmers and because you have more primer dimer molecules then your specific DNA, it results in higher efficiency. You can check that in melting curve analysis graph representing – (d/dt)fluorescence (483-533) as fluorescence fluctuation or 0.3 higher is than initial fluorescence from 60 °C. You can also try to measure fluorescence not at 72 C but instead at higher temp ( primer dimers dissociate normally around 80 C if the primers are not GC rich). So it can be safe to measure fluorescence 5 degrees lower than the melting temp of specific product Good LuckFollowing
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Answer added in Western Blot41 Trouble with large molecular weight extracellular matrix proteinsBy Daniella Pizzurro · University of Washington SeattleBehrouz Zandieh Doulabi · Academisch Centrum Tandheelkunde AmsterdamFor ECM proteins 3-8% Tris acetate based gels and runing buffer gives the best results for protein separation depending on size of gels (90 minutes ... [more]For ECM proteins 3-8% Tris acetate based gels and runing buffer gives the best results for protein separation depending on size of gels (90 minutes at 100-130V. Did you check if the problems lies in electropheriss (commasie blue staining of the gel) or on protein transfer to membrane (ponceu red stainnig of the blots)??Following
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Answer added in Western Blot5 Casein for blockingBy George Komis · Palacký University of OlomoucBehrouz Zandieh Doulabi · Academisch Centrum Tandheelkunde AmsterdamDear George You can use both low fat milk and or 0.5% Caseine for detection of phosoproteins on the western blots depending on detection method. Howev... [more]Dear George You can use both low fat milk and or 0.5% Caseine for detection of phosoproteins on the western blots depending on detection method. However, using milk at blocking reagent you should monitor pH because during overnight incubation with primary antibodies the lactic acid present in milk lowers the pH in both PBS and TBS based buffers. For more info see the links under http://www.pall.com/main/Laboratory/Literature-Library-Details.page?id=48923 http://www.thermoscientific.com/ecomm/servlet/productscatalog_11152__92506_-1_4 http://www.piercenet.com/browse.cfm?fldID=5A414328-5056-8A76-4EA5-51555CF0015D&utm_source=thermoscientific&utm_medium=node&utm_term=blocking-buffers-for-protein-methods&utm_campaign=value-prop-punchouts http://wolfson.huji.ac.il/purification/PDF/PhosphoProtein/PhosTag_PAGE_SDS.pdf Good luckFollowing
Publications (62) View all
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Article: Immunolocalization and western blotting of the anion exchanger pendrin in ameloblasts.
European Journal Of Oral Sciences 08/2012; 120(4):369-72. · 1.88 Impact Factor -
Article: Differentiation of human adipose-derived stem cells towards cardiomyocytes is facilitated by laminin.
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ABSTRACT: Adipose-derived stem cells (ASCs) are promising candidates for therapy in myocardial infarction (MI). However, the frequency of human ASCs that differentiate towards cardiomyocytes is low. We hypothesized that adherence to extracellular matrix molecules that are upregulated after MI might increase human stem cell differentiation towards cardiomyocytes. We analysed putative ASC differentiation on fibronectin-coated, laminin-coated and uncoated culture plates. Expression of cardiac markers in cells was analysed 1, 3 and 5 weeks after stimulation with 5-aza-2-deoxycytidine. After 1 week, mRNA expression of myosin light chain-2alpha (MLC-2alpha), an early marker in cardiomyocyte development, was increased significantly in treated cells, independent of coating. At 5 weeks, however, mRNA expression of the late cardiomyocyte development marker SERCA2alpha was only significantly increased in 5-aza-2-deoxycytidine-treated cells cultured on laminin. Significantly higher numbers of cells were immunopositive for MLC-2alpha in cultures of treated cells grown on laminin-coated wells, when compared with cultures of treated cells grown on uncoated wells, both at 1 week and at 5 weeks. Furthermore, after 3 weeks, significantly more alpha-actinin- and desmin-positive cells were detected after treatment with 5-aza-2-deoxycytidine, but only in uncoated wells. After 5 weeks, however, the number of desmin-positive cells was only significantly increased after treatment of cells with 5-aza-2-deoxycytidine and culture on laminin (61% positive cells). Thus, we have found that a high percentage of human ASCs can be differentiated towards cardiomyocytes; this effect can be improved by laminin, especially during late differentiation.Cell and Tissue Research 12/2008; 334(3):457-67. · 3.11 Impact Factor -
Article: Beta1 integrins regulate chondrogenesis and rock signaling in adipose stem cells.
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ABSTRACT: Beta1 integrins play a controversial role during chondrogenesis. Since the maturation of chondrocytes relies on a signaling switch from cell-cell to cell-matrix interactions, we hypothesized that beta1 integrins play a different role at the earlier (mainly cell-cell interaction) from the later stage (mainly cell-matrix interaction) of chondrogenesis. Our data showed: in plain medium, sox9, collagen X, and collagen II gene expressions of ASCs were induced by beta1-integrin blockage at day 14. In chondrogenic medium, however, sox 9, sox6, and collagen II gene expression were induced at day 4 but inhibited at day 14. In addition, both beta1-integrin blockage and TGF-beta1 down-regulated Rock-1 and -2 gene expression and produced the round cells. We concluded that beta1 integrins play a more important role at the later stages than earlier stages of chondrogenesis, and that the onset of chondrogenesis promoted by beta1-integrin blockage might be through inhibiting Rock signaling.Biochemical and Biophysical Research Communications 09/2008; 372(4):547-52. · 2.48 Impact Factor -
Article: The polymine spermine regulates osteogenic differentiation in adipose stem cells.
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ABSTRACT: For bone tissue engineering, it is important that mesenchymal stem cells (MSCs) differentiate into osteoblasts. To develop a method for differentiation of adipose tissue-derived mesenchymal stem cells (AT-MSCs) along the osteogenic lineage, we studied the effect of polyamines, which are organic cations implicated in bone growth and development, on differentiation of AT-MSCs. Treatment of goat-derived AT-MSCs with 1,25-dihydroxyvitamin-D3 (1,25(OH)(2)D(3)), which stimulates osteogenic differentiation, for 7 days induced gene expression of the polyamine-modulated transcription factor-1 (PMF-1) and spermidine/spermine N (1)-acetyltransferase (SSAT), which are both involved in polyamine metabolism, suggesting that polyamines are involved in osteogenic differentiation of AT-MSCs. Furthermore, treatment of AT-MSCs with the polyamine spermine-regulated gene expression of runx-2, a transcription factor involved in early stages of osteogenic differentiation, and that of osteopontin, a bone matrix protein expressed in later stages of osteogenic differentiation. Runx-2 gene expression was increased 4 and 14 days after a short 30 min. treatment with spermine, while osteopontin gene expression was only increased 4 days after spermine treatment. Finally, alkaline phosphatase activity, which is intimately involved in the formation of extracellular matrix of bone, was increased 4 weeks after the 30 min.-spermine treatment of AT-MSCs. In conclusion, this study shows for the first time that the polyamine spermine regulates differentiation of AT-MSCs along the osteogenic lineage, which can be used as a new method for differentiation of AT-MSCs along the osteogenic lineage. Therefore, polyamines may constitute a promising tool for bone tissue engineering approaches using AT-MSCs, such as a one-step surgical procedure for spinal interbody fusion.Journal of Cellular and Molecular Medicine 06/2008; 12(5A):1710-7. · 4.13 Impact Factor -
SourceAvailable from: Behrouz Zandieh Doulabi
Article: Influence of collagen type II and nucleus pulposus cells on aggregation and differentiation of adipose tissue-derived stem cells.
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ABSTRACT: Tissue microenvironment plays a critical role in guiding local stem cell differentiation. Within the intervertebral disc, collagen type II and nucleus pulposus (NP) cells are two major components. This study aimed to investigate how collagen type II and NP cells affect adipose tissue-derived stem cells (ASCs) in a 3D environment. ASCs were cultured in collagen type I or type II hydrogels alone, or co-cultured in transwells with micromass NP cells for 4 and 14 days. ASCs seeded in collagen type II gels acquired dentritic cell shapes, and orchestrated cell density-dependent gel contraction rates. Up-regulation of collagen type X, but not of other chondrogenic markers was observed at day 4, irrespective of the hydrogel type. Strikingly, in co-cultures with NP cells, more pronounced differentiation of ASCs along the cartilaginous lineage was observed (up-regulation of collagen IIA, IIB and aggrecan gene expression, as well as stronger alcian blue staining), when ASCs were embedded in collagen type II in comparison with type I hydrogels. Interestingly, strong cellular condensations/aggregations were observed in ASC-seeded type II, but not type I gels, and this aggregation was markedly delayed when the same gels were co-cultured with NP cells. The NP cell-mediated inhibition of ASC aggregation in collagen type II gels coincided with down-regulation of integrin subunit alpha2 gene expression. We conclude that soluble factors released by NP cells can direct chondrogenic differentiation of ASCs in collagen hydrogels, and that combination with a nucleus-mimicking collagen type II microenvironment enhances differentiation towards a more pronounced cartilage/NP lineage relative to collagen type I hydrogels.Journal of Cellular and Molecular Medicine 03/2008; 12(6B):2812-22. · 4.13 Impact Factor