您好,欢迎光临上海伟进生物科技有限公司
请登录 | 免费注册         服务热线:4006-800-892
  首页 > 公司新闻 > Systembio去外泌体血清Exosome-depleted FBS EXO-FBS-50A-1
今日推荐
总排行
Systembio去外泌体血清Exosome-depleted FBS EXO-FBS-50A-1
作者:汤姆     来源:伟进生物     发布日期:2017-05-03

Exosome-depleted FBS Media Supplement - USA Certified    50 ml       EXO-FBS-50A-1
SBI 去外泌体血清 现货促销 单独订购一瓶可获赠100元京东卡一张。
Exo-FBS Overview: When You're Not Studying Bovine Exosomes


Do you study bovine exosomes? If not, and you use fetal bovine serum (FBS) in your cell or tissue culture media, you may be isolating more bovine exosomes than you realize. Serum is a rich source of exosomes, FBS included. In order to help scientists study only the exosomes they want, SBI provides Exo-FBS™ exosome-depleted FBS.

  • Exosome-sized vesicles removed
  • Very low levels of CD63 positive cow exosomes
  • Undetectable levels of cow microRNAs
  • Comparable growth rates as standard FBS
  • Identical use as standard FBS (add 10% in DMEM or RPMI)
   

Background

Standard growth media for most cells in culture require FBS as a growth supplement to DMEM. FBS is derived from bovine (cow) serum and contains a high abundance of cow exosome vesicles. These exosomes can interfere or cause significant background issues when studying the exosomes secreted from your cells of interest in standard culture conditions.

SBI has developed an exosome-depleted FBS growth supplement called Exo-FBS that has been stripped of bovine exosomes. Exo-FBS supports equivalent growth of many types of cells in culture, is devoid of cow CD63-positive exosomes, and does not have any measurable bovine microRNAs. Cell growth in Exo-FBS is comparable to standard FBS. The process of manufacturing of Exo-FBS is a patented method in Patent No.: US 9,005,888 B2.

Conclusions

Perform your studies of cellular secreted exosomes in culture without the worry of contaminating cow exosomes in your experiments, and with no ultracentrifugation required when you use ExoQuick-TC. Available as standard FBS supplement or heat inactivated FBS media supplement (treated at 65°C for 15 minutes before bovine exosome removal).


Exo-FBS Has Greatly Reduced Levels of Bovine Exosomes

   

Use SBI’s Exo-FBS to ensure your exosome preparations from cell or tissue culture media contain only the exosomes from your cells, not from the FBS in your media. Both ELISA and NanoSight analysis demonstrate the great reduction in bovine exosome levels in Exo-FBS compared to standard FBS.

NanoSight Particle Analysis Show Low Exosome Levels in Exo-FBS

To demonstrate the depletion of exosomes in our Exo-FBS product, we diluted standard FBS and Exo-FBS samples 1:1000 and then analyzed for particle size and abundance using a NanoSight LM10 instrument. The standard FBS sample shows a significant amount of exosome-sized microvesicles where the Exo-FBS exosome-depleted FBS sample has a drastic reduction in exosome particles.

Exo-FBS is Depleted of Bovine CD63 Exosomes

   

The tetraspanin CD63 protein is a common marker for exosomes. We utilized a bovine-specific anti-CD63 antibody to develop an Enzyme Linked Immunosorbent Assay (ELISA). Equal volumes (50 µl) of either standard FBS or Exo-FBS depleted media supplement were used in this ELISA assay. Amounts of CD63-positive bovine exosomes are dramatically reduced in Exo-FBS compared to standard FBS (graphed results are normalized to the signal level of standard FBS).

   

   

Exo-FBS is Cleaner than Ultracentrifuged FBS

Exo-FBS is cleaner than UC FBS

Quality Control data is generated on every batch of Exo-FBS produced at SBI by comparing NanoSight particle count analyses to the source FBS, FBS ultracentrifuged for 18 hours and SBI's exosome-depleted Exo-FBS products. All samples were diluted 1:100 and NTA data collected in triplicate.

Exo-FBS Has Undetectable Levels of Bovine microRNAs


Not only does Exo-FBS have greatly reduced levels of exosomes, bovine microRNAs in Exo-FBS are undetectable as shown by highly sensitive qPCR assays. Standard FBS and Exo-FBS media supplements (4 ml) were treated with Trizol extraction methods to recover exosomal RNAs. RNA was converted to cDNA and 72 individual bovine microRNAs were measured by qPCR using SBI's QuantiMir system. Of the 72 microRNAs tested, 12 yielded amplification curves in the FBS sample but not in the Exo-FBS sample.

Exo-FBS Is Identical to Standard FBS for Cell Growth

Not only does Exo-FBS enable confident isolation of exosomes free from contaminants present in standard FBS, it supports identical growth rates as standard FBS. 

To compare growth rates of cells in Exo-FBS versus standard FBS, we grew HT1080 fibrosarcoma cells, PC-3 prostate cancer cells, MCF-7 breast cancer cells, and HEK293 cells in complete medium with either 10% standard FBS or 10% Exo-FBS supplement. Cells were seeded at either 10,000 or 20,000 cells, and then cultured under standard conditions at 37°C with 5% CO2 for 5 days in the medium indicated. The cells were imaged calculate growth rates and observe cellular morphologies. Equivalent growth and similar cellular morphologies were observed for standard FBS and Exo-FBS media tested across these 4 cell lines.

Online Resources

Product Citations

Connor Y, Tekleab S, Nandakumar S, Walls C, Tekleab Y, Husain A, Gadish O, Sabbisetti V, Kaushik S, Sehrawat S, Kulkarni A, Dvorak H, Zetter B, R Edelman E, Sengupta S. Physical nanoscale conduit-mediated communication between tumour cells and the endothelium modulates endothelial phenotype. Nat Commun. 2015 Dec 16;6:8671.

Almanza G, Zanetti M. High-efficiency Generation of Multiple Short Noncoding RNA in B-cells and B-cell-derived Extracellular Vesicles. Mol Ther Nucleic Acids. 2015 Dec 15;4:e271.

Sanchez CA, Andahur EI, Valenzuela R, Castellon EA, Fulla JA, Ramos CG, Trivino JC. Exosomes from bulk and stem cells from human prostate cancer have a differential microRNA content that contributes cooperatively over local and pre-metastatic niche. Oncotarget. 2015 Dec 9. doi: 10.18632/oncotarget.6540.

Holder BS, Jones T, Sancho Shimizu V, Rice TF, Donaldson B, Bouqueau M, Forbes K, Kampmann B. Macrophage exosomes induce placental inflammatory cytokines: a novel mode of maternal-placental messaging. Traffic. 2015 Nov 24. doi: 10.1111/tra.12352.

Hiroki Sakakura, Shinji Mii, Sumitaka Hagiwara, Takuya Kato, Noriyuki Yamamoto, Hideharu Hibi, Masahide Takahashi, Yoshiki Murakumo. CD109 is a component of exosome secreted from cultured cells. Biochemical and Biophysical Research Communications. 2015 doi:10.1016/j.bbrc.2015.12.063.

Zhang L, Zhang S, Yao J, Lowery FJ, Zhang Q, Huang WC, Li P, Li M, Wang X, Zhang C, Wang H, Ellis K, Cheerathodi M, McCarty JH, Palmieri D, Saunus J, Lakhani S, Huang S, Sahin AA, Aldape KD, Steeg PS, Yu D. Microenvironment-induced PTEN loss by exosomal microRNA primes brain metastasis outgrowth. Nature. 2015 Oct 19. doi: 10.1038/nature15376.

Philley JV, Kannan A, Qin W, Sauter ER, Ikebe M, Hertweck KL, Troyer DA, Semmes OJ, Dasgupta S. Complex-I alteration and enhanced mitochondrial fusion are associated with prostate cancer progression. J Cell Physiol. 2015 Nov 4. doi: 10.1002/jcp.25240.

Nakase I, Kobayashi NB, Takatani-Nakase T, Yoshida T. Active macropinocytosis induction by stimulation of epidermal growth factor receptor and oncogenic Ras expression potentiates cellular uptake efficacy of exosomes. Sci Rep. 2015 Jun 3;5:10300. doi: 10.1038/srep10300.

Khan MB, Lang MJ, Huang MB, Raymond A, Bond VC, Shiramizu B, Powell MD. Nef exosomes isolated from the plasma of individuals with HIV-associated dementia (HAD) can induce Aβ1-42 secretion in SH-SY5Y neural cells. J Neurovirol. 2015 Sep 25. [Epub ahead of print].

Nakase I, Futaki S. Combined treatment with a pH-sensitive fusogenic peptide and cationic lipids achieves enhanced cytosolic delivery of exosomes. Sci Rep. 2015 May 26;5:10112. doi: 10.1038/srep10112.

Momen-Heravi F, Bala S, Kodys K, Szabo G. Exosomes derived from alcohol-treated hepatocytes horizontally transfer liver specific miRNA-122 and sensitize monocytes to LPS. Sci Rep. 2015 May 14;5:9991. doi: 10.1038/srep09991.

Peterson MF, Otoc N, Sethi JK, Gupta A, Antes TJ. Integrated systems for exosome investigation. Methods. 2015 Apr 24. pii: S1046-2023(15)00161-9.

Zhang Y, Chopp M, Meng Y, Katakowski M, Xin H, Mahmood A, Xiong Y. Effect of exosomes derived from multipluripotent mesenchymal stromal cells on functional recovery and neurovascular plasticity in rats after traumatic brain injury. J Neurosurg. 2015 Jan 16:1-12.

Greening DW, Xu R, Ji H, Tauro BJ, Simpson RJ. A protocol for exosome isolation and characterization: evaluation of ultracentrifugation, density-gradient separation, and immunoaffinity capture methods. Methods Mol Biol. 2015;1295:179-209.

Jaideep S. and Gossett, Daniel R. and Tse, Henry T. K. and Lamm, Robert J. and Kulkarni, Rajan P. and Carlo, Dino Di.Rapid inertial solution exchange for enrichment and flow cytometric detection of microvesicles. Biomicrofluidics 9, 014112 (2015).

Josson S, Gururajan M, Sung SY, Hu P, Shao C, Zhau HE, Liu C, Lichterman J, Duan P, Li Q, Rogatko A, Posadas EM, Haga CL, Chung LW. Stromal fibroblast-derived miR-409 promotes epithelial-to-mesenchymal transition and prostate tumorigenesis. Oncogene. 2014 Jul 28;0. doi: 10.1038/onc.2014.212.

Bukong TN, Momen-Heravi F, Kodys K, Bala S, Szabo G. Exosomes from hepatitis C infected patients transmit HCV infection and contain replication competent viral RNA in complex with Ago2-miR122-HSP90. PLoS Pathog. 2014 Oct 2;10(10):e1004424.

Momen-Heravi F, Bala S, Bukong T, Szabo G. Exosome-mediated delivery of functionally active miRNA-155 inhibitor to macrophages. Nanomedicine. 2014 Mar 29. pii: S1549-9634(14)00132-4.

Basu S, Bhattacharyya SN. Insulin-like growth factor-1 prevents miR-122 production in neighbouring cells to curtail its intercellular transfer to ensure proliferation of human hepatoma cells. Nucleic Acids Res. 2014;42(11):7170-85.

Kaur S, Singh SP, Elkahloun AG, Wu W, Abu-Asab MS, Roberts DD. CD47-dependent immunomodulatory and angiogenic activities of extracellular vesicles produced by T cells. Matrix Biol. 2014 May 30. pii: S0945-053X(14)00092-4.

Vargas A, Zhou S, Ethier-Chiasson M, Flipo D, Lafond J, Gilbert C, Barbeau B. Syncytin proteins incorporated in placenta exosomes are important for cell uptake and show variation in abundance in serum exosomes from patients with preeclampsia. FASEB J. 2014 May 8. [Epub ahead of print].

Deschout H, Raemdonck K, Stremersch S, Maoddi P, Mernier G, Renaud P, Jiguet S, Hendrix A, Bracke M, Van den Broecke R, Roding M, Rudemo M, Demeester J, De Smedt SC, Strubbe F, Neyts K, Braeckmans K. On-chip light sheet illumination enables diagnostic size and concentration measurements of membrane vesicles in biofluids. Nanoscale. 2014 Jan 16;6(3):1741-7.

Ramteke A, Ting H, Agarwal C, Mateen S, Somasagara R, Hussain A, Graner M, Frederick B, Agarwal R, Deep G.Exosomes secreted under hypoxia enhance invasiveness and stemness of prostate cancer cells by targeting adherens junction molecules. Mol Carcinog. 2013 Dec 17. doi: 10.1002/mc.22124.

Xin H, Li Y, Cui Y, Yang JJ, Zhang ZG, Chopp M. Systemic administration of exosomes released from mesenchymal stromal cells promote functional recovery and neurovascular plasticity after stroke in rats. J Cereb Blood Flow Metab. Aug 2013, doi: 10.1038/jcbfm.2013.152.

Xin H, Li Y, Liu Z, Wang X, Shang X, Cui Y, Gang Zhang Z, Chopp M. Mir-133b Promotes Neural Plasticity and Functional Recovery after Treatment of Stroke with Multipotent Mesenchymal Stromal Cells in Rats Via Transfer of Exosome-Enriched Extracellular Particles. Stem Cells. 2013 Apr 30. doi: 10.1002/stem.1409.

Chen L, Wang Y, Pan Y, Zhang L, Shen C, Qin G, Ashraf M, Weintraub N, Ma G, Tang Y. Cardiac progenitor-derived exosomes protect ischemic myocardium from acute ischemia/reperfusion injury. Biochem Biophys Res Commun. 2013 Jan 11. pii: S0006-291X(13)00051-X. doi: 10.1016/j.bbrc.2013.01.015.

Technical References

Adachi T, Nakanishi M, Otsuka Y, Nishimura K, Hirokawa G, Goto Y, Nonogi H, Iwai N. Plasma microRNA 499 as a biomarker of acute myocardial infarction. Clin Chem. 2010 Jul;56(7):1183-5.

De Smaele E, Ferretti E, Gulino A. MicroRNAs as biomarkers for CNS cancer and other disorders. Brain Res. 2010 Jun 18;1338:100-11.

Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, Pogosova-Agadjanyan EL, Peterson A, Noteboom J, O'Briant KC, Allen A, Lin DW, Urban N, Drescher CW, Knudsen BS, Stirewalt DL, Gentleman R, Vessella RL, Nelson PS, Martin DB, Tewari M. Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A. 2008 Jul 29;105(30):10513-8.

Laterza OF, Lim L, Garrett-Engele PW, Vlasakova K, Muniappa N, Tanaka WK, Johnson JM, Sina JF, Fare TL, Sistare FD, Glaab WE. Plasma MicroRNAs as sensitive and specific biomarkers of tissue injury. Clin Chem. 2009 Nov;55(11):1977-83.

Valadi H, Ekström K, Bossios A, Sjöstrand M, Lee JJ, Lötvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol. 2007 Jun;9(6):654-9.

Pegtel DM, Cosmopoulos K, Thorley-Lawson DA, van Eijndhoven MA, Hopmans ES, Lindenberg JL, de Gruijl TD, Wordinger T, Middeldorp JM. Functional delivery of viral miRNAs via exosomes. Proc Natl Acad Sci USA; 2010 Apr 6; 107(14):6328-33.

Mathivanan, S. and Simpson, R.J. ExoCarta: A compendium of exosomal proteins and RNA. Proteomics. 2009.21, 4997-5000.

Thery C, Ostrowski M, Segura E. Membrane vesicles as conveyors of immune responses. Nat Rev Immunol. 2009. 8, 581-93.

Michael A, Bajracharya SD, Yuen PS, Zhou H, Star RA, Illei GG, Alevizos I. Exosomes from human saliva as a source of microRNA biomarkers. Oral Dis; 2010 Jan; 16(1):34-8.

Luo SS, Ishibashi O, Ishikawa G, Ishikawa T, Katayama A, Mishima T, Takizawa T, Shigihara T, Goto T, Izumi A, Ohkuchi A, Matsubara S, Takeshita T, Takizawa T. Human villous trophoblasts express and secrete placenta-specific microRNAs into maternal circulation via exosomes. Biol Reprod; 2009 Oct; 81(4):717-29.

Taylor DD, Gercel-Taylor C. MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. Gynecol Oncol; 2008 Jul; 110(1):13-21.

Simpson RJ, Lim JW, Moritz RL, Mathivanan S. Exosomes: proteomic insights and diagnostic potential. Expert Rev Proteomics. 2009 Jun;6(3):267-83. Review.

新手指南
配送方式
关注伟进公众号
扫一扫免费电话咨询
QQ在线咨询/订购:314870818       电话:021-54960076    18930085161
地址:上海市 徐汇区 喜泰路239号 汇丰创意园 8号302B      传真:021-33685277     电子邮件:wegene_china@126.com
版权所有©2017 上海伟进生物科技有限公司 泸ICP备17008735号