Abmgood不朽系列商品 (immortalized cell)-BV-2 Cells

BV-2 細胞 (貨號T9077)

BV-2 細胞特性

細胞型態: Adherent, microglial。 來源Brain。解凍後，細胞需要4-5天才能恢復培養。 細胞培養條件: PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture.  Dulbecco’s Modified Eagle Medium (DMEM), High Glucose (TM500) + 10% FBS(Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂.

Cat. No.	T9077
Name	BV-2 Cells
Description	Microglia cell line derived from C57BL/6 mouse.
Organism	Mouse (M. musculus)
Tissue	Brain
Donor History	Female, 1 week, C57BL/6
Growth Properties	Adherent, microglial
Cell Type	Immortalized Cells
Storage Condition	Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions	Ship with dry ice.
Product Format	Frozen
Intended Use	This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
BioSafety	II
Certificate of Analysis	For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions	PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture.  Dulbecco’s Modified Eagle Medium (DMEM), High Glucose (TM500) + 10% FBS(Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. *Do not heat-inactivate

HOT!! 黴漿菌PCR偵測 | DAPI染核 | 細胞內黴漿菌去除劑 | 水浴槽抑菌劑 | 噴霧式去黴劑 等系列商品

Abmgood不朽化系列商品 (immortalized reagenet) Ready-to-use

Abmgood公司於人類與老鼠不朽之細胞株上，具有幾十年的實驗經驗；提供一系列不朽 (immortalized)化之細胞株、及不朽化之試劑商品。ready-to-use定量好之retrovirus、lentiviru、 adenovirus，可直接加於細胞中進行感染表現；病毒液包括有SV40重組病毒、EBV重組病毒、hTERT病毒液、Myc基因病毒液、p53基因病毒液、Rb基因病毒液、Ras基因病毒液。提供106及109兩種包裝力價之病毒液；亦提供一系列病毒力價濃縮與定量試劑商品。

細胞不朽化的工具中，SV40 T antigen是最簡單且最常用來造成不朽化細胞生成的方法。多數的病毒基因和抑癌基因（p53、Rb等）的不活化相關，會誘導細胞處於replicative senescence的階段。目前也有研究顯示被感染的細胞，SV40 T anbtigen會誘導端粒酶（Telomerase）的活性。與端粒（telomere）長度相關的Telomerase Reverse Transcriptase protein (TERT)是一種不朽細胞的研究應用。平常細胞中，hTERT是處於不活化態，必要時會進行活化保持telomere的長度，避免細胞發生replicative senescence的現象。某些細胞hTERT 過度表現（over-expression）對於誘發細胞不朽化是無效的（如初代上皮細胞），反而會造成細胞死亡；同時表現hTERT catalytic subunit以及p53或RB siRNA在人類初代卵巢上皮細胞會造成細胞不朽化。過度表現Ras或Myc T58A mutants也有初代細胞不朽化的現象產生。

Abmgood不朽化系列商品 (immortalized cell)

根據細胞類型，Primary cells初代細胞在體外具有小於10-50次傳代的有限壽命。 不朽化細胞株能夠延長增殖，並且具有與親代組織相似或相同的基因型和表現型。

哺乳動物細胞的不朽化Immortalized策略

存在用於在培養物中不朽化哺乳動物細胞的幾種方法。 Abmgood多年研發經驗，開發出了一系列Immortalized商品；包括即用型逆轉錄病毒，慢病毒和腺病毒載體。

Gene Overexpressed	Viruses Offered	Application
hTERT	Adenovirus, Lentivirus, Retrovirus	可靠和常用。 可能在一些細胞類型（例如上皮細胞）中引起細胞死亡
SV40 T Antigen	Adenovirus, Lentivirus, Retrovirus	可靠和常用。 通常用於不朽化上皮細胞。
EBV genes	Epstein-Barr Virus	不朽化B淋巴細胞，有時是T淋巴細胞。
HPV E6-E7	Lentivirus	不朽化角化細胞。
HOXB8, HOXA9, HOXA10	Lentivirus	不朽化造血細胞，包括巨噬細胞，造血祖細胞和骨髓祖細胞。
Adenoviral E1A-E1B	Lentivirus	不朽化大範圍的初級囓齒動物細胞，包括羊水細胞，原發性上皮細胞，成纖維細胞和原代腎細胞。
p53, Rb siRNA	Lentivirus	與hTERT共表達以更好地使上皮細胞不朽化。
Bmi1	Lentivirus	不朽化上皮細胞。
CDK4	Lentivirus	與hTERT共同表達更真實的細胞模型。
Myc T58A (c-Myc), Ras V12 (H-Ras)	Lentivirus	不朽化各種細胞，包括神經幹細胞，初代腎細胞，成紅細胞和各種上皮細胞（原發性前列腺，乳腺，氣道等）。

 

Thawing細胞解凍方法：(每種細胞建議條件可能有許不同，依照原廠建議操作，以下為通用方式) 操作方式下載

1.在37℃水浴中解凍小瓶，直到只有一小塊冰塊。(DMSO對細胞有毒性，over-thaw過度解凍下可能造Stress)

2.將細胞轉移到含有5ml pre-warmed complete media 之15ml管中。

3.將細胞以290xg離心5分鐘。

4.小心丟棄上清液，不要打擾細胞沉澱，輕輕地上下移液，將細胞重新懸浮在1ml完全培養基中。

5.將細胞培養在50,000 – 100,000 cells/cm²。 cells will have viability of around 40-50%. Post thawing, it will need 4-5 days for cells to recover in culture; Recommended split ratio is 1:4 to 1:5

DMSO在細胞凍存中的應用

二甲基亞碸是一種重要的滲透型細胞保護劑。在深低溫（零下200度）保存細胞時凍存過程中，為防止細胞內液冰晶形成、滲透壓改變、細胞結構紊亂等導致的損傷，有必要使用含有DMSO冷凍保護劑。DMSO能夠快速穿透細胞膜進入細胞中，降低冰點、延緩凍存過程，同時提高細胞內離子濃度，減少細胞內冰晶的形成，從而減少細胞損傷。深低溫時二甲基亞碸的細胞毒性受到抑制。復蘇時動作要快，儘快洗掉二甲基亞碸，否則會造成對細胞嚴重的毒性。二甲基亞碸（DMSO）是目前最好的細胞凍存保護劑，但也是一種以細胞毒性很大的化學試驗劑。研究結果表明，培養液中DMSO濃度為10%時，細胞生長抑制率近100%；1‰濃度時抑制率為35%，即使是0.04‰的濃度，DMSO對細胞的生長也有不利的影響。

Subculture細胞繼代方法: (每種細胞建議條件可能有許不同，依照原廠建議操作，以下為通用方式) 操作方式下載

1.當細胞70-90％ confluency時，建議繼代培養。

2. Aspirate old media。懸浮液中的一些細胞仍然是活細胞。或者，也可以收集含有懸浮細胞的上清液並離心（Skip to Step 6 in protocol）。

3.加入1：1比例的1X無菌PBS和0.25％胰蛋白酶-EDTA（TM051）。

4.在室溫下孵育細胞3-5分鐘並攪拌培養容器直至90％的細胞脫落。

5. 中和胰蛋白酶:添加等量之trypsin + PBS。

6.收集細胞並以290xg離心5分鐘。

7.丟棄上清液，輕輕地上下移液，將細胞重新懸浮在完全培養基中。

8.建議split ratio比不超過1：3。

Freezing細胞凍結：(每種細胞建議條件可能有許不同，依照原廠建議操作，以下為通用方式) 操作方式下載

Complete growth medium with decomplemented FBS to a final concentration of 50% from PAN Biotech (P40-37500) or from Nucleus Biologics (FBS1824-001) and 5% DMSO.

儲存溫度：液氮氣相。重要提示：此細胞係對FBS批次非常敏感。我們強烈建議最終用戶使用從PAN biotech或Nucleus Biologics 之FBS。

慢病毒包裝系列產品Lentivirus慢病毒低速純化(純化) | Lentivirus 慢病毒高濃度純化(純化) | Lentivirus病毒RT-QPCR定量(定量) 

Cat No	Product Name
T0016	Immortalized Human Cord Blood Mesenchymal Stromal Cells (cbMSC-hTERT)
T0017	Immortalized Human Cord Blood Mesenchymal Stromal Cells with RFP (cbMSC-hTERT-RFP)
T0533	Immortalized Human Endometrial Stromal Cells (HESC)
T0697	Immortalized Human Cranial Suture Progenitor Cells (iSuPs)
T0723	Immortalized Human Foreskin Keratinocytes (HFK-16E6E7)
T0727	Immortalized Human Prostate Stromal Cells (PM151T) – hTERT
T0752	Immortalized Human Osteoarthritis Cartilage Mesenchymal Stem Cells (OASC5)
T0764	Immortalized Human Endometriotic Cell Line (12Z)
T0841	Immortalized Human Dental Pulp Stem Cells (TP-023)
T0842	Immortalized Human Dental Pulp Stem Cells (TP-037)
T1013	Immortalized Human Diploid Fibroblasts – hTERT
T1015	Immortalized Human Lung Fibroblast – hTERT
T9435	Immortalized Human Cancer Associated Fibroblast Cells – SV40
T3000	Acan CRISPR Knockout RCS Cell Line (Rat)
T3002	Aquaporin-4 Stable 293 Cell Line
T3005	MuSK Stable HEK293 Cell Line
T3006	Luciferase Stable PC3 Cell Line
T3008	Luciferase (Luc2) Stable TRAMP Cell Line
T3009	Luciferase Stable U87MG/ΔEGFR Cell Line
T3013	Luciferase Stable WM-266-4 Cell Line
T3015	NF-kB Luciferase Stable RAW264.7 Cell Line
T3016	HEK293-CymR-Puro Stable Cell Line
T3017	A2058-CymR Stable Cell Line
T3018	SKMEL5-CymR Stable Cell Line
T3021	GRPR-Luciferase Stable PC3 Cell Line
T3024	Bax Knockout Immortalized Mouse Kidney Epithelial Cell Line
T3025	Bak Knockout Immortalized Mouse Kidney Epithelial Cell Line
T3026	Bax/Bak Double Knockout Immortalized Mouse Kidney Epithelial Cell Line
T3027	Atg5 Knockout Immortalized Mouse Kidney Epithelial Cell Line
T3028	Atg7 Knockout Immortalized Mouse Kidney Epithelial Cell Line
T3030	Nav2.1/β1/Contactin Stable Chinese Hamster Lung Cell Line
T3031	PARP-1 Knockout Immortalized Mouse Heart Endothelial (HYKO6) Cell Line
T3032	Dual Notch1-YFP and Inducible Delta-mCherry Reporter CHO-K1 (hN1) Cell Line
T3033	Dual Notch1(Variant)-YFP and Inducible Delta-mCherry Reporter CHO-K1 (hN1G4<sup>esn</sup>) Cell Line
T3034	TLR3 Knockout Immortalized Mouse Dendritic Cell Line
T3035	TLR9 Knockout Immortalized Mouse Dendritic Cell Line
T3036	Type I IFN Receptor Knockout Immortalized Mouse Dendritic Cell Line
T3037	Ryanodine Receptor 1 Stable HEK293 Cell Line
T3038	KCC2 Stable HEK293 Cell Line
T3039	Stable HEK293 Expressing Mutant UQCRB-MT1 Cell Line
T3040	c-Kit/D816V Stable Ba/F3 Cell Line
T3041	c-Kit/WT Stable Ba/F3 (with GNNK- isoform) Cell Line
T3042	c-Kit/WT Stable Ba/F3 (with GNNK+ isoform) Cell Line
T3043	c-Kit/Y568F Stable Ba/F3 Cell Line
T3044	c-Kit/Y570F Stable Ba/F3 Cell Line
T3045	c-Kit Y568F/Y570F Stable Ba/F3 Cell Line
T3046	c-Kit/Y703F Stable Ba/F3 Cell Line
T3047	c-Kit/Y936F Stable Ba/F3 Cell Line
T3048	c-Kit Y703F/Y936F Stable Ba/F3 Cell Line
T3049	c-Kit/N705AStable Ba/F3 Cell Line
T3050	c-Kit/N938A Stable Ba/F3 Cell Line
T3051	c-Kit N705A/N938A Stable Ba/F3 Cell Line
T3052	c-Kit/D816H Stable Ba/F3 Cell Line
T3053	c-Kit/D816F Stable Ba/F3 Cell Line
T3054	c-Kit/D816YStable Ba/F3 Cell Line
T3055	c-Kit/V560D Stable Ba/F3 Cell Line
T3056	c-Kit/K642E Stable Ba/F3 Cell Line
T3057	c-Kit/Y721F Stable Ba/F3 Cell Line
T3058	c-Kit/Y823F Stable Ba/F3 Cell Line
T3059	TLR2 Stable HEK293 Cell Line
T3060	PGC-1α/ERRα Stable 293T Cell Line
T3061	TNK1 Stable Ba/F3 Cell Line
T3062	ALK Stable Ba/F3 Cell Line
T3063	LTK Stable Ba/F3 Cell Line
T3064	AXL Stable Ba/F3 Cell Line
T3065	MERTK Stable Ba/F3 Cell Line
T3066	TYRO3 Stable Ba/F3 Cell Line
T3067	EPHA1 Stable Ba/F3 Cell Line
T3068	EPHA5 Stable Ba/F3 Cell Line
T3069	EPHA7 Stable Ba/F3 Cell Line
T3070	EPHA8 Stable Ba/F3 Cell Line
T3071	FGFR2 Stable Ba/F3 Cell Line
T3072	FGFR3 Stable Ba/F3 Cell Line
T3073	IGF1R Stable Ba/F3 Cell Line
T3074	INSR Stable Ba/F3 Cell Line
T3075	JAK1 Stable Ba/F3 Cell Line
T3076	JAK2 Stable Ba/F3 Cell Line
T3077	JAK3 Stable Ba/F3 Cell Line
T3078	MET Stable Ba/F3 Cell Line
T3079	MST1R Stable Ba/F3 Cell Line
T3080	FLT3 Stable Ba/F3 Cell Line
T3081	KIT Stable Ba/F3 Cell Line
T3082	PDGFRA Stable Ba/F3 Cell Line
T3083	PDGFRB Stable Ba/F3 Cell Line
T3084	RET Stable Ba/F3 Cell Line
T3085	ROS1 Stable Ba/F3 Cell Line
T3086	BLK Stable Ba/F3 Cell Line
T3087	FGR Stable Ba/F3 Cell Line
T3088	FYN Stable Ba/F3 Cell Line
T3089	HCK Stable Ba/F3 Cell Line
T3090	LCK Stable Ba/F3 Cell Line
T3091	SRC Stable Ba/F3 Cell Line
T3092	YES1 Stable Ba/F3 Cell Line
T3093	SYK Stable Ba/F3 Cell Line
T3094	NTRK1 Stable Ba/F3 Cell Line
T3095	NTRK2 Stable Ba/F3 Cell Line
T3096	NTRK3 Stable Ba/F3 Cell Line
T3097	FLT4 Stable Ba/F3 Cell Line
T3098	ERBB2 Stable Ba/F3 Cell Line
T3099	EPHA4 Stable Ba/F3 Cell Line
T3100	ABL1 Stable Ba/F3 Cell Line
T3101	ABL2 Stable Ba/F3 Cell Line
T3102	AZ-AHR Stable HepG2 Luciferase Reporter Cell Line
T3103	AZ-GR Stable HeLa Luciferase Reporter Cell Line
T3104	AIZ-AR Stable 22Rv1 Luciferase Reporter Cell Line
T3105	BMP Responsive Immortalized Reporter (BRITER) Cell Line
T3106	EGFR Stable Ba/F3 Cell Line
T3107	ERBB4 Stable Ba/F3 Cell Line
T3108	EPHA2 Stable Ba/F3 Cell Line
T3109	EPHB1 Stable Ba/F3 Cell Line
T3110	EPHB2 Stable Ba/F3 Cell Line
T3111	TYK2 Stable Ba/F3 Cell Line
T3112	EPHB4 Stable Ba/F3 Cell Line
T3113	FER Stable Ba/F3 Cell Line
T3115	LYN Stable Ba/F3 Cell Line
T3116	ZAP70 Stable Ba/F3 Cell Line
T3117	TIE1 Stable Ba/F3 Cell Line
T3118	KDR Stable Ba/F3 Cell Line (IL-3 Independent)
T3119	TNK2 Stable Ba/F3 Cell Line
T3120	CSK Stable Ba/F3 Cell Line
T3121	MATK Stable Ba/F3 Cell Line
T3122	DDR1 Stable Ba/F3 Cell Line
T3123	DDR2 Stable Ba/F3 Cell Line
T3124	PTK2 Stable Ba/F3 Cell Line
T3126	EPHA10 Stable Ba/F3 Cell Line
T3127	EPHA3 Stable Ba/F3 Cell Line
T3128	EPHA6 Stable Ba/F3 Cell Line
T3129	EPHB3 Stable Ba/F3 Cell Line
T3131	PTK2B Stable Ba/F3 Cell Line
T3132	FGFR4 Stable Ba/F3 Cell Line
T3133	INSRR Stable Ba/F3 Cell Line
T3134	AATK Stable Ba/F3 Cell Line
T3136	LMTK3 Stable Ba/F3 Cell Line
T3137	MuSK Stable Ba/F3 Cell Line
T3138	CSF1R Stable Ba/F3 Cell Line
T3139	PTK7 Stable Ba/F3 Cell Line
T3140	ROR1 Stable Ba/F3 Cell Line
T3141	ROR2 Stable Ba/F3 Cell Line
T3142	RYK Stable Ba/F3 Cell Line
T3143	FRK Stable Ba/F3 Cell Line
T3145	SRMS Stable Ba/F3 Cell Line
T3146	BMX Stable Ba/F3 Cell Line
T3147	BTK Stable Ba/F3 Cell Line
T3148	ITK Stable Ba/F3 Cell Line
T3149	TEC Stable Ba/F3 Cell Line
T3150	TXK Stable Ba/F3 Cell Line
T3151	TEK2 Stable Ba/F3 Cell Line
T3152	FLT1 Stable Ba/F3 Cell Line
T3153	STYK1 Stable Ba/F3 Cell Line
T3154	SPINK1 Stable AsPC-1 Cell Line
T3155	hSSAO/VAP-1 A7r5 Stable Cell Line
T3156	Mouse R-spondin1 Stable 293T Cell Line
T3157	Human T Lymphoblast Stably Deoxycytidine Kinase Deficient (AraC-8C) Cell Line
T3158	TrkA Stable T17 Cell Line
T3159	TrkB Stable T48 Cell Line
T3160	Luc-Akt-PH Stable MCF7 Cell Line
T3161	Luc-Akt-PH/YFP Stable MCF-7/B2 Stable Cell Line
T3162	HEK293S-TetR Stably Expressing TP Cell Line
T3163	HEK293S-TetR GnTI- Stably Expressing TP Cell Line
T3164	HEK293S-TetR Stably Expressing A160T Cell Line
T3165	HEK293S-TetR GnTI- Stably Expressing A160T Cell Line
T3166	Cx26 Stable BxPC-3 Cell Line
T3167	E-cadherin Stable MIA PaCa-2 Cell Line
T3168	BMP9 Stable HEK293 Cell Line
T3169	92 TAg Stable Y-LIZ Wildtype Mouse Embryonic Fibroblast Cell Line
T3170	88 Tag Stable Y-LIZ DNA Polymerase-β Knockout Mouse Embryonic Fibroblast Cell Line
T3171	SIRT1 Wiltype Stable RKO Cell Line
T3172	SIRT1 Knockout Stable RKO Cell Line
T3173	SIRT1 Wildtype Stable DLD-1 Cell Line
T3174	SIRT1 Knockout Stable DLD-1 Cell Line
T3175	SIRT1 Wildtype Stable 293T Cell Line
T3176	SIRT1 Knockout Stable 293T Cell Line
T3177	PZ-TR Stable HepG2 Luciferase Reporter Cell Line
T3178	Ad5 pTP Stable 293 (293pTP) Cell Line
T3181	p47phox Stable Knockout Mouse Aortic Endothelial (iMAEC-p47) Cell Line
T3182	Podoplanin Stably Expressing CHO Cell Line
T3183	Stable Retinal Pigment Epithelial Expressing Truncated hELG1-CFP Cell Line
T3184	Stable EGFR and Erbb2 Conditional Knockdown Mouse Skin Keratinocyte (B1B2F3) Cell Line
T3185	Stable Erbb2 Conditional Knockdown Mouse Skin Keratinocyte (B2F6) Cell Line
T3186	Stable EGFR Conditional Knockdown Mouse Skin Keratinocyte (Egfr2) Cell Line
T3187	Stable Mouse Myeloid-derived Suppressor-like LAL Knock Out (HD1B) Cell Line
T3188	Stable Rat Expressing HA-tagged SCL26A9 Thyroid Epithelial (SLC26A9-FRT) Cell Line
T3189	Stable VERO Overexpressing HER2 Cell Line
T3190	Stable VERO Overexpressing HER3 Cell Line
T3191	Stable VERO Overexpressing HER2 and HER3 Cell Line
T3192	Stable VERO Overexpressing HER1 Cell Line
T3193	Stable CHO Overexpressing HER2 Cell Line
T3194	Stable VERO Overexpressing HER1 and HER2 Cell Line
T3195	Stable HepG2 Expressing Murine ADH1 (VA-13) Cell Line
T3196	Stable HA-Tagged hGALE Expressing CHO Cell Line
T3197	Stable G90E Mutated hGALE Expressing CHO Cell Line
T3198	Stable V94M Mutated hGALE Expressing CHO Cell Line
T3199	Stable C307Y Mutated hGALE Expressing CHO Cell Line
T3203	CRE Expressing 293 Cell Line
T3207	ICP47 Stable Raji Cell Line
T3208	ICP47 Stable Mel JuSo Cell Line
T3209	HLA-DQ8 Expressing Stable K562 Cell Line
T3210	Mutant AKT1 Stable Proteus Syndrome Derived Cell Line (F6B)
T3217	GFP Stably Expressing Schwann cells
T3218	huGRPr Expessing Stable Ace-1 Cell Line
T3219	Apolipoprotein A-1 Expressing Stable MC38 Cell Line (MC38 Apo)
T3220	Apolipoprotein A-1 and P144 Over-expressing Stable MC38 Cell Line (MC38 ApoLinkerP144)
T3222	BRCA1 L1780P Mutant Stable HCC1937 Cell Line
T3223	BRCA1 L1780P Mutant Stable HCC1395 TNBC Cell Line
T3224	NDUFS2 Stable Knockout HEK293 Cell Line (HEK293ΔNDUFS2)
T3229	HEK293-T Cells Stably Expressing MLL-AF6 (MA6)
T3230	HEK293-T Cells Stably Expressing AF6-MLL (A6M)
T3231	HEK293-T Cells Stably Expressing MLL-AF6/AF6-MLL (CO1)
T3232	HEK293-T Cells Stably Expressing exMLL-AF6 (exMA6)
T3233	HEK293-T Cells Stably Expressing AF6-shMLL (A6shM)
T3234	HEK293-T Cells Stably Expressing exMLL-AF6/AF6-shMLL (CO2)
T3235	A549 Cells Stably Expressing hACE2 Receptor and TMPRSS2 (A549-AT)
T3236	U2OS Stably Expressing NONO-APEX2 Cell Line (U2OS:NONO-APEX2-HA)
T3241	ts-46 Stable Knockout HEK293 Cell Line (ts-46 293KO)
T3242	ts-101 Stable Knockout HEK293 Cell Line (ts-101 293KO)
T3243	Human Progerin Expressing 3T3-L1 Stable Cell Line (3T3L1-PG)