hESC BG01V (RMCL-00260321)
Cat. No.: RMCL-00260321
Category: Cell Lines
INQUIRY
1 vial
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 Features
| BSL | BSL-1 |
|---|---|
| Product Format | Frozen |
| Applications | Stem cell research |
| Specific Applications | This product is pluripotent and can differentiate to representatives of the three primary germ layers. |
| Product Type | Human cells |
| Organism | Homo sapiens, human |
| Tissue | Embryo; Inner cell mass |
| Morphology | Spherical colony |
| Cell Type | Embryonic Stem Cell |
| 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 |
| Karyotype | 49, XXY, +12, +17 |
| Antigen Expression | The cells stain positive for pluripotency markers and alkaline phosphatase activity. |
QC
| Mycoplasma Contamination | Not detected |
|---|
Storage & Shipping
| Storage Conditions | Vapor phase of liquid nitrogen |
|---|---|
| Unpacking and Storage Instructions | Check all containers for leakage or breakage. 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. |
| Temperature | 37°C |
| Atmosphere | 95% Air, 5% CO2 |
| 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. 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 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). 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. 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. Spin the cells at 270 x g for 5 minutes. Aspirate the supernatant and resuspend the pellet in 3 ml of complete medium. 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. 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. |
For research use only. Not for clinical use.
