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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 运输方式：
冻存运输
- ATCC Number：
SCRC-2002™
- 组织来源：
inner cell mass
- 物种来源：
人
- 是否是肿瘤细胞：
0
- 细胞形态：
球形
- 库存：
大量
- 细胞类型：
胚胎干细胞
- 器官来源：
胚胎
- 年限：
embryo, blastocyst
| Designations: | hESC BG01V | ||
| Depositors: | BresaGen, Inc. | ||
| Biosafety Level: | 1 | ||
| Shipped: | frozen | ||
| Medium & Serum: | See Propagation | ||
| Organism: | Homo sapiens | ||
| Morphology: | spherical colony |
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| Source: | Organ: embryo Tissue: inner cell mass Cell Type: embryonic stem cell; |
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| Permits/Forms: | In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location. | ||
| Restrictions: | Prior to purchase, for-profit commercial institutions must obtain a license agreement. For instructions on how to proceed, please contact the ATCC Office of Licensing and Business Development at licensing@ATCC .org or 703 365 2773. | ||
| Applications: | ATCC SCRC-1040.2 or ATCC SCRC-1040.1 ) exhibit uniform morphology, a predictable growth rate, and are easy to maintain in culture. BG01V cells are pluripotent and can differentiate to representatives of the three primary germ layers. BG01V was derived from the wild-type, parental hESC line BG01 [PMID 12968106, PMID 15153607]. The cells stain positive for pluripotency markers and alkaline phosphatase activity. |
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| Cytogenetic Analysis: | 49, XXY, +12, +17 | ||
| Age: | embryo, blastocyst | ||
| Comments: | BG01V is a human embryonic stem cell line with an abnormal karyotype. BG01V was derived from the wild-type, parental hESC line BG01 [PMID 12968106, PMID 15153607]. Despite the abnormal karyotype, BG01V cell colonies grown on irradiated or Mitomycin C-treated Mouse embrynic fibroblast (MEFs - e.g. ATCC SCRC-1040.2 or ATCC SCRC-1040.1 ) exhibit uniform morphology, a predictable growth rate, and are easy to maintain in culture. BG01V cells are pluripotent and can differentiate to representatives of the three primary germ layers. The cells stain positive for pluripotency markers and alkaline phosphatase activity. | ||
| Propagation: | ATCC complete growth medium: 1:1 Mixture of Dulbecco's Modified Eagles Medium and Ham's F-12 medium containing 1.2 g/L sodium bicarbonate, 2.5 mM L-glutamine, 15 mM HEPES and 0.5 mM sodium pyruvate (ATCC 30-2006) supplemented with 2.0 mM L-Alanyl-L-Glutamine (ATCC 30-2115), 0.1 mM Non-essential amino acids (ATCC 30-2116), 0.1 mM 2-mercaptoethanol (Sigma Catalog No. M-7522) and 4 ng/ml bFGF (R& D Systems Catalog No. 233-FB), 80%; Knockout serum replacement (Invitrogen Catalog No. 10828), 5%; fetal bovine serum (ATCC SCRR-30-2020), 15% Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C |
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| Subculturing: | Protocol: To insure the highest level of viability, be sure to warm media to 37C before using it on the cells. The passaging ratio depends on the density/confluency of the colonies. It ranges between 1:3 and 1:6. Note: If the colonies are close to or touching each other the culture is overgrown (refer to images on the SCC website - stemcells.atcc.org). Overgrowth will result in differentiation. 1. At least 24 hours prior to each passage, plate treated MEFs onto the culture vessels to be used. Base the number of dishs/flasks to be used on the passaging ratio (see product sheet). 2. Prepare 0.5 mg/ml or approximately 200 units/ml Collagenase IV solution (Invitrogen 17104-019) in DMEM/F12 and sterile filter using 0.22 micrometer low-protein binding filter. Check the units/mg for each lot of powder. 3. Remove medium from cells. Add appropriate volume (see product sheet) of Collagenase IV solution. 4. Incubate at 37C for up to 2 hours. 5. Check the cells after the first 30 minutes and then every 15 minutes. When the majority of the hESC colonies have completely detached or the edges of the colonies have rounded up, add appropriate amount (see product sheet) of DMEM/F12 and wash gently using a pipette. Under optimal conditions, all the colonies can be washed off with feeder cells left behind. If some colonies are still attached, gently scrape the surface area with the tip of a 5 ml pipette if necessary. 6. Collect cell suspensions into a 50 ml conical tube. 7. Centrifuge for 5 minutes at 200 x g at 25C. 8. Remove the supernatant and resuspend in complete growth medium. Pipette up and down to break the colonies to smaller clumps and evenly distribute cells to feeder-covered dishes/flasks. 9. Add complete growth medium to each tissue culture vessel to achieve the appropriate final volume. Please consult the product sheet for details. Medium Renewal: Every day after the first 48 hours |
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| Preservation: | Freeze medium: culture medium, 70%; fetal bovine serum, 20%; DMSO, 10% Storage temperature: liquid nitrogen vapor phase |
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| References: | 90326: Zeng X, et al. Properties of pluripotent human embryonic stem cells BG01 and BG02. Stem Cells 22: 292-312, 2004. PubMed: 15153607 | ||
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文献和实验相关实验
Procedure for Culturing BG01V Human Embryonic Stem Cells with Human Feeder Cell Conditioned Medium
with the BG01V line of hES cells (3, 4). Please note that other hES cell lines may require modifications of this protocol. Optimal culture conditions must be determined by the investigator for each hES line. Materials Required Reagents:
Culturing BG01V Human Embryonic Stem Cells with Mouse Embryonic Fibroblast (MEF)-Conditioned Media
If culturing in the absence of a feeder cell layer is desired, human embryonic stem (hES) cells can be maintained using Mouse or Human-Conditioned Media (Catalog # AR005 , AR007 ). The protocol below has been used with the BG01V line
Q：Annexin V 凋亡检测试剂盒的原理是什么？ A：Annexin V 是一种钙离子依赖性磷脂结合蛋白，与磷脂酰丝氨酸 PS 有高度亲和力，可通过细胞外侧暴露的 PS 与凋亡早期细胞胞膜结合，将 Annexin V 标记荧光染料如 FITC 利用流式细胞仪或荧光显微镜可检测细胞凋亡。 碘化丙啶(Propidium Iodide, PI)或 7-AAD 可与双链 DNA 特异性结合，并产生强烈的荧光，正常情况下无法透过细胞膜。由于凋亡晚期或坏死细胞膜丧失完整性，核染料 PI 或 7-AAD
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hESC BG01V
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