在哺乳動物的大腦中，絕大部分細(xì)胞是星形膠質(zhì)細(xì)胞。它們形態(tài)各異，并與神經(jīng)元各個(gè)部分緊密相連，因此可能具有多種功能。星形膠質(zhì)細(xì)胞為神經(jīng)元提供結(jié)構(gòu)，營養(yǎng)和代謝上的支持并調(diào)節(jié)突觸活動。此外，星形膠質(zhì)細(xì)胞可提示許多神經(jīng)疾病的病理過程。腦卒中及其他神經(jīng)損傷后，正常星形膠質(zhì)細(xì)胞功能受損，可嚴(yán)重影響神經(jīng)元的存活。腦損傷后的長期恢復(fù)，通過軸突生長，突觸可塑性，或神經(jīng)元再生，也影響星形膠質(zhì)細(xì)胞表面分子的表達(dá)和營養(yǎng)因子釋放。大量的研究已經(jīng)表明，星形膠質(zhì)細(xì)胞是在**神經(jīng)系統(tǒng)中的*功能多樣化的細(xì)胞群。在體外研究中，星形膠質(zhì)細(xì)胞的培養(yǎng)為探索細(xì)胞多樣性提供了重要工具。
ScienCell實(shí)驗(yàn)室的HA提取自大腦皮層，原代凍存，冰凍運(yùn)輸。每管細(xì)胞密度*過1×106/ml。細(xì)胞經(jīng)GFAP免疫熒光鑒定。本細(xì)胞經(jīng)檢測不含HSC?HIV-1,?HBV,?HCV,?支原體,?細(xì)菌,?酵母菌和真菌。如采用ScienCell?實(shí)驗(yàn)室特制的培養(yǎng)基，可保證此細(xì)胞10次倍增。
推薦培養(yǎng)基：星形膠質(zhì)細(xì)胞培養(yǎng)基(AM, Cat. #1801)
|
貨號
|
1800
|
|
產(chǎn)地
|
美國
|
|
縮寫
|
HA
|
|
規(guī)格
|
5 x 10^5 /1ml
|
|
用途
|
科研
|
|
儲存
|
液氮
|
|
運(yùn)輸
|
干冰
|
1.) Zhou Y, Hayashi I, Wong J, Tugusheva K, Renger JJ, Zerbinatti C. (2014) 'Intracellular Clusterin Interacts with Brain Isoforms of the Bridging Integrator 1 and with the Microtubule-Associated Protein Tau in Alzheimer's Disease.'
2.) Monticone M, Taherian R, Stigliani S, Carra E, Monteghirfo S, Longo L, Daga A, Dono M, Giaretti W, Castagnola P. (2014) 'NAC, Tiron and Trolox Impair Survival of Cell Cultures Containing Glioblastoma Tumorigenic Initiating Cells by Inhibition of Cell Cycle Progression.'
3.) Cruz-Orengo L, Daniels BP, Dorsey D, Basak SA, Grajales-Reyes JG, McCandless EE, Piccio L, Schmidt RE, Cross AH, Crosby SD, Klein RS. (2014) 'Enhanced sphingosine-1-phosphate receptor 2 expression underlies female CNS autoimmunity susceptibility.' J clin invest.
4.) Cruz-Orengo L, Daniels BP, Dorsey D, Basak SA, Grajales-Reyes JG, McCandless EE, Piccio L, Schmidt RE, Cross AH, Crosby SD, Klein RS. (2014) "Enhanced sphingosine-1-phosphate receptor 2 expression underlies female CNS autoimmunity susceptibility." J clin invest. 124: 2571-84.
5.) Monticone M, Taherian R, Stigliani S, Carra E, Monteghirfo S, Longo L, Daga A, Dono M, Giaretti W, Castagnola P. (2014) "NAC, Tiron and Trolox Impair Survival of Cell Cultures Containing Glioblastoma Tumorigenic Initiating Cells by Inhibition of Cell Cycle Progression." PLOS ONE. 9: e90085.
6.) Zhou Y, Hayashi I, Wong J, Tugusheva K, Renger JJ, Zerbinatti C. (2014) "Intracellular Clusterin Interacts with Brain Isoforms of the Bridging Integrator 1 and with the Microtubule-Associated Protein Tau in Alzheimer's Disease." PloS one. 9: e103187.
7.) Urich E, Patsch C, Aigner S, Graf M, Iacone R, Freskgrd P. (2013) "Multicellular self-assembled spheroidal model of the blood brain barrier." Sci Rep. 3: 1500.
8.) Abdelmoaty S, Wigerblad G, Bas DB, Codeluppi S, Fernandez-Zafra T, El-Awady ES, Moustafa Y, Abdelhamid AE, Brodin E, Svensson CI. (2013) "Spinal Actions of Lipoxin A4 and 17(R)-Resolvin D1 Attenuate Inflammation-Induced Mechanical Hypersensitivity and Spinal TNF Release." PLoS One. 8: e75543.
9.) Cucullo L, Hossain M, Tierney W, Janigro D. (2013) "A new dynamic in vitro modular capillaries-venules modular system: Cerebrovascular physiology in a box." BMC Neuroscience. 14:18
10.) Vijayaraghavan S, Karami A, Aeinehband S, Behbahani H, Grandien A, Nilsson B, Ekdahl KN, Lindblom RP, Piehl F, Darreh-Shori T. (2013) "Regulated Extracellular Choline Acetyltransferase Activity - The Plausible Missing Link of the Distant Action of Acetylcholine in the Cholinergic Anti-Inflammatory Pathway." PLoS One. 8: e65936
11.) Li J, Bonifati S, Hristov G, Marttila T, Valmary Degano S, Stanzel S, Schnolzer M, Mougin C, Aprahamian M, Grekova SP, Raykov Z, Rommelaere J, Marchini A. (2013) 'Synergistic combination of valproic acid and oncolytic parvovirus H?1PV as a potential therapy against cervical and pancreatic carcinomas.'
12.) Arscott W, Tandle AT, Zhao S, Shabason JE, Gordon IK, Schlaff CD, Zhang G, Tofilon PJ, Camphausen KA. (2013) 'Ionizing Radiation and Glioblastoma Exosomes: Implications in Tumor Biology and Cell Migration.'
13.) Arscott W, Tandle AT, Zhao S, Shabason JE, Gordon IK, Schlaff CD, Zhang G, Tofilon PJ, Camphausen KA. (2013) "Ionizing Radiation and Glioblastoma Exosomes: Implications in Tumor Biology and Cell Migration." Transloncol. 6: 638-48.
14.) Li J, Bonifati S, Hristov G, Marttila T, Valmary?Degano S, Stanzel S, Schnolzer M, Mougin C, Aprahamian M, Grekova SP, Raykov Z, Rommelaere J, Marchini A. (2013) "Synergistic combination of valproic acid and oncolytic parvovirus H?1PV as a potential therapy against cervical and pancreatic carcinomas." EMBO Mol Med. 5: 1537-55.
15.) Corti S, Nizzardo M, Simone C, Falcone M, Donadoni C, Salani S, Rizzo F, Nardini M, Riboldi G, Magri F, Zanetta C, Faravelli I,Bresolin N, Comi GP. (2012) "Direct reprogramming of human astrocytes into neural stem cells and neurons." Exp Cell Res. 318: 1528-41.
16.) Muller FL, Colla S, Aquilanti E, Manzo VE, Genovese G, Lee J, Eisenson D, Narurkar R, Deng P, Nezi L, Lee MA, Hu B, Hu J, Sahin E, Ong D, Fletcher-Sananikone E, Ho D, Kwong L, Brennan C, Wang YA, Chin L, DePinho RA. (2012) "Passenger deletions generate therapeutic vulnerabilities in cancer." Nature. 2488: 337-42
17.) Triscott J, Lee C, Hu K, Fotovati A, Berns R, Pambid M, Luk M, Kast RE, Kong E, Toyota E, Yip S, Toyota B, Dunn SE. (2012) "Disulfiram, a drug widely used to control alcoholism, suppresses the self-renewal of glioblastoma and over-rides resistance to temozolomide." Oncotarget. 3: 1112-23.
18.) Cucullo L, Hossain M, Puvenna V, Marchi N, Janigro D. (2011) "The role of shear stress in Blood-Brain Barrier endothelial physiology." BMC Neurosci. 12: 40.
19.) Ghosh C, Gonzalez-Martinez J, Hossain M, Cucullo L, Fazio V, Janigro D, Marchi N. (2010) "Pattern of P450 expression at the human blood-brain barrier: roles of epileptic condition and laminar flow." Epilepsia. 51: 1408-17.
20.) Das A, Banik NL, Ray SK. (2010) "Flavonoids activated caspases for apoptosis in human glioblastoma T98G and U87MG cells but not in human normal astrocytes." Cancer. 116: 164-76.
21.) Furr SR, Moerdyk-Schauwecker M, Grdzelishvili VZ, Marriott I. (2010) "RIG-I mediates nonsegmented negative-sense RNA virus-induced inflammatory immune responses of primary human astrocytes." Glia. 58: 1620-9.
22.) Koob AO, Paulino AD, Masliah E. (2010) "GFAP reactivity, apolipoprotein E redistribution and cholesterol reduction in human astrocytes treated with alpha-synuclein." Neurosci Lett. 469: 11-4.
23.) Ruiz S, Brennand K, Panopoulos AD, Herrer as A, Gage FH, Izpisua-Belmonte JC. (2010) "High-efficient generation of induced pluripotent stem cells from human astrocytes." PLoS One. 5: e15526.
24.) Sanchez MC, Benitez A, Ortloff L, Green LM. (2009) "Alterations in glutamate uptake in NT2-derived neurons and astrocytes after exposure to gamma radiation." Radiat Res. 171: 41-52.
25.) Williams R, Dhillon NK, Hegde ST, Yao H, Peng F, Callen S, Chebloune Y, Davis RL, Buch SJ. (2009) "Proinflammatory cytokines and HIV-1 synergistically enhance CXCL10 expression in human astrocytes." Glia. 57: 734-43.
26.) Williams R, Yao H, Dhillon NK, Buch SJ. (2009) "HIV-1 Tat co-operates with IFN-gamma and TNF-alpha to increase CXCL10 in human astrocytes." PLoS One. 4: e5709.
27.) Cucullo L, Couraud P, Wekslet B, Romero I, Hossain M, Rapp E, Janigro D. (2008) "Immortalized human brain endothelial cells and flow-based vascular modeling: a marriage of convenience for rational neurovascular studies." J Cereb Blood Flow Metab. 28: 312-28.
28.) Hoelzinger DB, Nakada M, Demuth T, Rosensteel T, Reavie LB, Berens ME. (2008) "Autotaxin: a secreted autocrine/paracrine factor that promotes glioma invasion." J Neurooncol. 86: 297-309.
29.) Qiu J, Ai L, Ramachandran C, Yao B, Gopalakrishnan S, Fields CR, Delmas AL, Dyer LM, Melnick SJ, Yachnis AT, Schwartz PH, Fine HA, Brown KD, Robertson KD. (2008) "Invasion suppressor cystatin E/M (CST6): high-level cell type-specific expression in normal brain and epigenetic silencing in gliomas." Lab Invest. 88: 910-25.
30.) Cucullo L, Hossain M, Rapp E, Manders T, Marchi N, Janigro D. (2007) "Development of a humanized in vitro blood-brain barrier model to screen for brain penetration of antiepileptic drugs." Epilepsia. 48: 505-16
31.) van den Pol AN, Robek MD, Ghosh PK, Ozduman K, Bandi P, Whim MD, Wollmann G. (2007) "Cytomegalovirus induces interferon-stimulated gene expression and is attenuated by interferon in the developing brain." J Virol. 81: 332-48.
32.) Konnikova L, Kotecki M, Kruger MM, Cochran BH. (2003) "Knockdown of STAT3 expression by RNAi induces apoptosis in astrocytoma cells." BMC Cancer. 3: 23.
?
1、Question:
Which genes could represent good housekeeping genes for RT-qPCR analysis of these cells? Any suggestions?
Answers:
We recommend β-actin, GAPDH, LDHA, NONO, and PPIH to serve as housekeeping genes for astrocyte qRT-PCR. We also recommend using multiple housekeeping genes for data normalization. In addition, in a qRT-PCR assay, we recommend using Genomic DNA (gDNA) Control (GDC) to detect possible gDNA contamination, and Positive PCR Control (PPC) to test whether samples contain inhibitors or other factors that may negatively affect gene expression results. ScienCell offers GeneQuery? Human Astrocyte Cell Biology qPCR Array Kit for astrocyte qRT-PCR analysis.
2、Question:
WAre this cell lines？
Answers:
They are primary cells, not immortalized cell lines. thanks for your interest
3、Question:
What is the recommended maximum number of passages that the primary human astrocytes can be grown to without losing phenotype? Is there a recommended freezing protocol?
Answers:
Human Astrocytes can be subcultured for 4-5 passages. However, for primary cells, the markers such as GFAP may be lost during each passage, even though they are still astrocytes. Therefore, we recommend that customers use the cells at early passage without expanding or passaging too much. We do not recommend re-freezing the cells for future use. thanks for your interest
