American Type Culture Collection (ATCC) Logo American Type Culture Collection (ATCC) Logo 0
  • Quick Order
  • Careers
  • Support

hESC BG01V

SCRC-2002

hESC BG01V is a fibroblast cell that was isolated from the inner cell mass of an embryo. hESC BG01V cells are pluripotent and can differentiate into representatives of the three primary germ layers.
Product category
Human cells
Organism
Homo sapiens, human
Cell type
embryonic stem cell
Morphology
spherical colony
Tissue
Embryo; Inner cell mass
Applications
Stem cell research
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
Buy Now
Price: $1,144.00 EA
Discounts may be available for our fellow nonprofit organizations. Login to see your price.

Generally ships within 1-3 business days

Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Handle as a potentially biohazardous material using universal precautions.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Detailed product information

General

Specific applications
BG01V cells are pluripotent and can differentiate to representatives of the three primary germ layers.

Characteristics

Derivation
BG01V is a human embryonic stem cell line with an abnormal karyotype. BG01V was derived from the wild-type, parental hESC line BG01. (Mitalipova M, Calhoun J, Shin S, Wininger D, Schulz T, Noggle S, Venable A, Lyons I, Robins A, Stice S. (2003). Human embryonic stem cell lines derived from discarded embryos. Stem Cells, 21(5), 521-6. PubMed: 12968106) (Zeng X, et al. Properties of pluripotent human embryonic stem cells BG01 and BG02. Stem Cells 22: 292-312, 2004. PubMed: 15153607)
Karyotype
49, XXY, +12, +17
Antigen expression
The cells stain positive for pluripotency markers and alkaline phosphatase activity.
Comments
BG01V is a human embryonic stem cell line with an abnormal karyotype.  Despite the abnormal karyotype, when grown on murine embryonic feeders (MEFs) these colonies exhibit uniform morphology, a predictable growth rate, and are easy to maintain in culture. The cells stain positive for pluripotency markers and alkaline phosphatase activity.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete 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%
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Feeder cells
Feeder cells may be grown in medium containing fewer growth factors than those required by the ES cells. Feeder cells are available from ATCC. Consult the product detail page and the product sheet provided for the feeder cells you wish to use for medium requirements. Feeder cells should be initiated 24 to 48 hours prior to inoculating with embryonic stem (ES) cells.
Handling procedure

To insure the highest level of viability thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at –70°C.  Storage at –70°C will result in loss of viability.

To insure the highest level of viability, be sure to warm media to 37ºC before using it on the cells. The cells were frozen in clumps since wild type Human ES cells experience low viability when dissociated to single cells.

  1. Plate radiation treated mouse embryonic fibroblasts (MEF, SCRC-1040.1) as a feeder layer onto appropriate size flask at least one day before thawing the vial. Use Table 1 to determine the correct density of feeders to plate (see product sheet for SCRC-10401.1 for protocol). One hour before thawing the vial of ES cells, perform a 100% medium change using complete growth medium (see below for recipe).  

Table 1. Plating Densities for MEFs

Flask/Plate

Growth Area (cm²)

CF-1 MEFs

T225

225

18.0 x 106

T75

75

7.0 x 106

T25

25

2.5 x 106

6 well

9.5

0.5 x 106

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 90 seconds).
  2. Remove the vial from the water bath before the cells are completely thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial’s contents plus 4 mL of complete growth medium to a 15 mL centrifuge tube. Use an additional 1 mL of media to rinse the vial and transfer the liquid to the 15 mL tube. Add 4 ml of complete growth medium to bring the total volume to 10 mL.
  4. Spin the cells at 270 x g for 5 minutes.
  5. Aspirate the supernatant and resuspend the pellet in 3 ml of complete medium.
  6. Add the 3 mL of cell suspension to one T25 flask or 3 wells of a 6 well plate (1 mL/well) containing feeders and growth medium.
  7. Incubate the culture at 37°C in a humidified 5% CO2/95% air incubator.

Do not change the medium for the first 48 hours. However, add an additional 4 ng/mL of bFGF 24 hours after the thaw. After the first 48 hours, change the medium daily.

Examine the colonies daily using an inverted microscope. It can take up to one week for colonies to appear. The first passage should occur 3-4 days after colonies are visible.
Subculturing procedure

To insure the highest level of viability, be sure to warm media to 37°C 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. 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.  Refer to Table 1 to determine the correct plating density for the feeders.
  2. Prepare 0.5 mg/mL or ~200 units/mL Collagenase IV solution (Invitrogen 17104-019) in DMEM/F12 and sterile filter using 0.22 µm low-protein binding filter. Check the units/mg for each lot of powder.
  3. Remove medium from cells. Add appropriate volume of Collagenase IV solution. Refer to Table 2 to determine the correct amount.
  4. Incubate at 37°C 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 of DMEM/F12 (Table 2) 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 25°C.
  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. Refer to Table #2 to find the appropriate volume based on surface area.

Table 2. Reagent Quantities

Flask/Plate

Growth Area (cm²)

Collagenase (ml)

DMEM/F12  (ml)

Growth Medium (ml)

T225

225

10

10

30

T75

75

3.0

5

12

T25

25

2

5

6

6 well

9.5

0.5

1

3

Medium Renewal

Every day after the first 48 hours

Complete Growth Medium for Feeder Cells

The feeder cells are grown in DMEM (ATCC #30-2002) supplemented with 15% FBS (ATCC # 30-2020).

Cryopreservation
  1. Follow the Subculturing Procedure above and use Collagenase IV to dissociate the cells.
  2. Centrifuge the cell suspension for 5 minutes at 200xg at 25°C.
  3. Resuspend the pellet in a 1:1 solution of 50% complete growth medium and 50% FBS. The total volume should be 0.5 ml times the number of vials to be frozen. Determine the number of vials using Table 3.
  4. Pipette up and down to break the colonies into small clumps. P1000 tips are used to efficiently break up the colonies.
  5. Slowly add an equal volume of complete growth medium with 20% DMSO. Mix gently.
  6. Evenly distribute 1 mL of the cell suspension into each cryovial.
  7. Store the vials in Styrofoam boxes at -80°C. Transfer the vials to liquid nitrogen 24 hours later.

 Table 3. # of Vials at 80-90% confluency

Flask/Plate

Growth Area (cm²)

# of vials

T225

225

16

T75

75

5

T25

25

2

6 well

9.5

1


Cryoprotectant Medium: Complete growth medium supplemented with 20% FBS and 10% DMSO. Follow the two step process in the Cryopreservation protocol. Cell-culture tested DMSO is available as ATCC Catalog No. 4-X.

Quality control specifications

Mycoplasma contamination
Not detected
STR profiling
Amelogenin: X,Y
CSF1PO: 10
D13S317: 11,12
D16S539: 9,11
D5S818: 10,12
D7S820: 10,11
TH01: 7,9.3
TPOX: 8
vWA: 16,17
D3S1358: 15,17
D21S11: 28,29
D18S51: 19
Penta_E: 13,14
Penta_D: 12
D8S1179: 10,12
FGA: 22
D19S433: 13,15
D2S1338: 17,24

History

Depositors
BresaGen, Inc.
Special collection
NSCR (National Stem Cell Resource)

Legal disclaimers

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.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

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. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

For-profit Research Use License from Viacyte

For-profit Organizations: For every order of this item, you must work directly with the contributor, Viacyte, acquired by Vertex, to (i) negotiate a research-use license, and/or (ii) have the contributor provide authorization to ATCC to ship this item under your existing license. We cannot ship this item until we receive communication directly from Vertex that we are authorized to ship each order.

We are providing the following contact information, but this information may change without notice:
Vertex
Attn: Steven Murfin
Email: [email protected] AND [email protected]

Once ATCC has received authorization from Vertex, your order will be reviewed, and this item will be released for shipment if all requirements are met. If you need assistance with your order, please contact our Customer Care team or your applicable distributor.

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Zeng X, et al. Properties of pluripotent human embryonic stem cells BG01 and BG02. Stem Cells 22: 292-312, 2004. PubMed: 15153607

Brimble S, et al. Karyotypic stability, genotyping, differentiation, feeder free maintenance and gene expression sampling in three human embryonic stem cell lines derived prior to August 9th 2001. Stem Cells Dev. 13(6): 585-597, 2004. PubMed: 15684826

Mitalipova M, Calhoun J, Shin S, Wininger D, Schulz T, Noggle S, Venable A, Lyons I, Robins A, Stice S. (2003). Human embryonic stem cell lines derived from discarded embryos. Stem Cells, 21(5), 521-6. PubMed: 12968106

Schulz TC, Noggle S, Palmarini G, Weiler D, Lyons I, Pensa K, Meedeniya A, Davidson B, Lambert N, Condie B (2004). Differentiation of Human Embryonic Stem Cells to Dopaminergic Neurons in Serum Free Suspension Culture. Stem Cells, 22(7), PMID: 15579641

Hay RJ, Caputo JL, Macy, ML, Eds. (1992) ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

View All Curated Citations for this Product

For product-related inquiries and issues, contact Technical Service:

Message Us

Hours of Operation

Monday - Friday
9:00am - 5:00pm
US Eastern Time