Dilute the thawed Cell Basement Membrane Gel to 150 µg/mL in cold DMEM: F-12 Medium ( ATCC 30-2006) by directly adding the Cell Basement Membrane Gel to the medium on ice and mix well.Thaw Cell Basement Membrane Gel in the refrigerator (2☌ to 8☌), on ice, overnight.(4 mL)(0.15 mg/mL) = 0.043 mL (14 mg/mL) Add 43 µL Cell Basement Membrane Gel directly into 4 mL DMEM: F12 To coat two 6 cm dishes, prepare as follows: Dilute Cell Basement Membrane in DMEM:F12 at a working concentration of 150 µg/mL: Protein concentration of Cell Basement Membrane (on product label): 14 mg/mL. Calculate the appropriate Cell Basement Membrane volume per plate based on concentration and usage. The concentration of Cell Basement Membrane is found on the product label.Įxample: 2 mL of Cell Basement Membrane Gel at 150 µg/mL is required to coat one 6 cm dish. Important: Cell Basement Membrane Gel will solidify in 15 to 30 minutes above 15° C. Keep Cell Basement Membrane Gel and labware on ice at all times to prevent the matrix from gelling prematurely. However, the colony morphology will recover after subsequent media change without ROCK inhibitor. The use of ROCK inhibitor may cause a transient spindle-like morphology effect on the cells. Note: Addition of ROCK inhibitor has been shown to increase the survival rate during subcultivation and thawing of human iPSCs. Stem cell culture medium with ROCK inhibitor must be used immediately. If using ROCK Inhibitor Y27632, prepare stem cell culture medium supplemented with final concentration of 10 μM ROCK Inhibitor Y27632. 30 Minutes Prior to Handling Cells – Pre-warm Pluripotent Stem Cell SFM XF/FF (stem cell culture medium) at 37☌ for at least 30 minutes before adding to cells.One Hour Prior to Thawing the iPS Cells – Prepare coated plates as described.Night before thawing iPSC cells – Thaw Cell Basement Membrane Gel on ice in refrigerator or cold room (2☌ to 8☌).Storage at –80☌ will result in loss of viability. If, upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at –80☌. To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. Stem Cell Dissociation Reagent ( ATCC ACS-3010) The base medium for this cell line is Pluripotent Stem Cell SFM XF/FF ( ATCC ACS-3002) which is a ready-to-use medium for serum-free and feeder-free iPSC culture. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below -130☌, preferably in liquid nitrogen vapor, until ready for use.ĪTCC iPSCs have been adapted to feeder- and serum-free culture conditions.Check all containers for leakage or breakage.Since then, it has received $4 million in equity financing from private investors. OncoCyte was formed in early 2009 to develop genetically modified stem cells capable of finding malignant tumors while carrying genes that can cause the destruction of the cancer cells. The firm will initially provide this technology to its majority-owned subsidiary, OncoCyte, for R&D related to genetically modified hESC-derived vascular progenitors designed to target and destroy malignant tumors. The phage display peptide technology licensed from SBMRI holds promise for use in directing human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) to sites in the body where they can have therapeutic effect, BioTime explains. By coating these peptides onto the surfaces of therapeutic cells using techniques that do not modify the cell physiology, Cell Targeting has produced tissue-specific and disease-specific cell modification agents with potential to take cell therapy products to a new level of performance, BioTime says. The technology acquired from CTI uses peptides selected for their ability to adhere to diseased tissues.
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