年限：embryo, blastocyst
器官來源：胚胎
細(xì)胞類型：胚胎干細(xì)胞
數(shù)量：大量
細(xì)胞形態(tài)：球形
是否是腫瘤細(xì)胞：0
物種來源：人
組織來源：inner cell mass
ATCC Number：SCRC-2002?
運(yùn)輸方式：凍存運(yùn)輸
規(guī)格：0.2ml Designations: hESC BG01V
Depositors: ?BresaGen, Inc.
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Organism: Homo sapiens
Morphology:spherical colony
Source: Organ: embryo
Tissue: inner cell mass
Cell Type: embryonic stem cell;
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.
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
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
Preservation: Freeze medium: culture medium, 70%; fetal bovine serum, 20%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
References: 90326: Zeng X, et al. Properties of pluripotent human embryonic stem cells BG01 and BG02. Stem Cells 22: 292-312, 2004. PubMed: 15153607
