STEM CELL GROWTH MEDIA
HIDEF-B8

HiDef-B8 400X Supplement Cat # LSS-204

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Defined, feeder-free maintenance medium supplement for human iPS cells. HiDef-B8 400X Supplement (1.25 mL), when added to 500 mL basal media (NOT INCLUDED) , is a complete, serum-free, defined formulation designed for the feeder-free maintenance and expansion of human induced pluripotent stem cells (iPSC) in the undifferentiated state. The HiDef-B8 400X Supplement is a unique formulation that offers modern conveniences of a flexible feeding schedule (including weekend-free maintenance) and also the ability to choose the matrix and passaging reagent that best suits specific applications.

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Defined Bioscience’s HiDef-B8 400X Supplement, when added to your basal media of choice (NOT INCLUDED), is a complete, serum-free, defined formulation designed for the feeder-free maintenance and expansion of human induced pluripotent stem cells (iPSC) in the undifferentiated state. The HiDef-B8 400X Supplement is a unique formulation that offers modern conveniences of a flexible feeding schedule (including weekend-free maintenance) and also the ability to choose the matrix and passaging reagent that best suits specific applications. HiDef-B8 contains insulin, ascorbic acid-2-phosphate (AA2P), transferrin, sodium selenite, thermostable FGF2/bFGF (aka FGF2-STAB or FGF2-G3), TGFB3, and NRG1. The addition of further components is not required. HiDef-B8 has been validated with DMEM/F12 from Corning® (Corning 10-092-CM) and may be used with either Matrigel (Corning) or Cultrex/Geltrex (Trevigen/Gibco) at concentrations as low as 2.5 µg cm-2 (a 1:800 dilution) or higher. Basal DMEM/F12 media and matrices from other manufactures may also be used once validated in your hands. Each lot of HiDef-B8 400X Supplement is used to prepare complete iPSC medium and then performance-tested in a culture assay using human iPSCs.

HiDef-B8 500X Supplement Cat
HiDef-B8 500X Supplement Cat

Highly optimized stem cell supplement for weekend-free cell maintenance

HiDef-B8 Stem Cell Medium Supplement is the formulation of a hiPSC culture medium that resulted from an exhaustive optimization of medium constituents and concentrations, establishing the necessity and relative contributions of each component to the pluripotent state and cell proliferation. The eight key ingredients are Insulin, Ascorbic acid 2-phosphate, Transferrin, Sodium selenite, FGF2-G3, TGFB3, NRG1, and Sodium bicarbonate. A recent publication from Northwestern University demonstrates the derivation and culture of 34 hiPSC lines in HiDef-B8 as well as the maintenance of pluripotency long term (over 100 passages). This formula also allows a weekend-free feeding schedule without sacrificing capacity for differentiation.

QUALIFICATION OF HIDEF-B8

The gold-standard demonstration of the suitability of a hiPSC medium is the capacity to generate hiPSC lines and maintain them in long-term culture. Our standard hiPSC line 19c3 has been cultured for >100 passages (p) in HiDEF-B8 and maintained expression of markers of undifferentiated status (SSEA4 and TRA-1-60) by flow cytometry (p131 at time of assay). We also generated hiPSC lines from 37 patients using established protocols but using B8. These hiPSC lines maintained SSEA4 and TRA-1-60 expression in culture (up to p65 at time of assay). In B8 hiPSCs maintained an hESC-like morphology; positive immunofluorescent staining for SSEA4, POU5F1, SOX2, and TRA-1-60; and normal karyotype.

Demonstration of the expression of markers of undifferentiated status in hiPSC line 19c3 cultured in B8 from passage (p) 21 to p131 (left) and 37 individual hiPSC lines derived in B8 from p12 to p65 (right), assessed by flow cytometry.

Phase-contrast images (original magnification 103) of hiPSC line 19c3 p44 cultured in B8. Scale bar, 100 mm.

Expression of markers of undifferentiated status in a variety of B8-derived hiPSC lines. Scale bar, 100 mm.

Example G-banding karyotype analysis of four hiPSC lines derived in B8.