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BMEM Human Primed Pluripotent Kit

BMEM Human Primed Pluripotent Kit

Optimized maintenance medium for human iPSC

Catalog #1001

Ships at room temperature

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$200.00 USD
$200.00 USD
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BMEM Human Primed Pluripotent, comprising of BMEM Basal Medium and Primed Supplement, is a highly optimized, chemically defined, animal origin-free (AOF) basal medium and supplement combination for the maintenance and expansion of human induced pluripotent stem cells (hiPSCs) in adherent culture.

BMEM Basal Medium is based on the rigorously optimized published formulation1.

Primed Supplement is new supplement specifically optimized for compatibility with adaptation of hiPSCs from legacy media such as mTeSR1™, a common problem with chemically defined media. Primed Supplement is optimized for component stability and to allow room temperature shipment.

Culture of hiPSCs in BMEM Human Primed Pluripotent results in increased cell proliferation and upregulation of core primed pluripotency factors when compared to legacy formulae1. BMEM Human Primed Pluripotent is formulated for compatibility with weekend-free and no-change feeding schedules protocols.

BMEM Human Primed Pluripotent is compatible with a variety of culture matrices, including Corning® Matrigel® GFR, Gibco™ Geltrex™ GFR, R&D Systems™ Cultrex™ UltiMatrix GFR, SigmaAldrich® ECMatrix™-511 E8 Laminin, Gibco™ Recombinant Vitronectin, Corning® Synthemax II-SC. 

BMEM Human Primed Pluripotent is compatible with a variety of passaging methods including BMEM Dissociation (1002), Gibco™ TrypLE™, Stemcell Technologies™ ReLeSR™, and Gibco™ Versene.

Each lot of BMEM Basal Medium and Primed Supplement is performance tested on hiPSC and tested for sterility.

Larger unit sizes are available on request.

Contains HEPES.

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Information Sheet Safety Data Sheet Quality Certificate

Product Information

The following components are sold as part of the BMEM Primed Pluripotent kit and are not available separately.
Component Name Size Storage
Primed Supplement Lyophilized powder, 2 mL vial Store at -20°C
BMEM Basal Medium 500 mL or 1000 mL Store at 4°C

Instructions/Directions for Use

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Instructions for Use

Use sterile technique to prepare complete BMEM Human Primed Pluripotent (basal medium + supplement). Complete BMEM can be used straight from the refrigerator (4 °C) and should not be warmed to RT or 37 °C prior to use. 

1. Add 1 mL of BMEM Basal Medium to the vial of Primed Supplement, pipette up and down gently until the powder becomes in solution (~5-10 times). Transfer the solution to the BMEM Basal Medium bottle and wash out the vial with additional BMEM Basal Medium. Gently mix the bottle by inversion.

2. Store complete BMEM at 4 °C for up to 30 days.

ROCK Inhibitor

Addition of a ROCK inhibitor, such as 10 µM Y27632, for 24 hours after passage is recommended to provide more reproducible passaging, reduce selective pressure, and allow higher split ratios. 

Incubator

hiPSC cultured in BMEM Human Primed Pluripotent should be maintained in a cell culture incubator at 37 °C with a humidified atmosphere of 5% CO2. Hypoxia (5% O2) can be used but is not essential.

Directions for Use

Daily media change

For this schedule, media should be changed daily and cells passaged every 4 days when they reach 70-80% confluence. When cells are adapted to BMEM, some lines will be able to be passaged at 1:20 or higher split ratios every four days. 

NOTE: If cells do not reach 70-80% confluence in five days then they should be passaged anyway with a variety of lower split ratios (i.e. 1:6, 1:12, 1:15). Do not allow cells to become >90% confluent as this can result in slow growth and unreliable passaging. 

NOTE: Not all cell lines will adapt to high split ratios such as 1:20 every four days, and if the wells are passaged at high split rations when they are too sparse (below 50% confluency), selective pressure will affect the cells, and spontaneous differentiation or other growth problems might arise.

Weekend-Free media change

BMEM Human Primed Pluripotent is compatible with the following weekend-free protocol:

  1. Passage the cells on Monday morning, seeding the cells in BMEM with ROCK inhibitor.
  2. Change the media on Tuesday morning adding 3x the volume of media (e.g., 6 mL per well in a 6 well plate).
  3. Skip media change on Wednesday.
  4. Passage cells on Thursday afternoon/evening, seeding the cells in BMEM with ROCK inhibitor.
  5. Change media on Friday afternoon/evening with 3x the volume of media.
  6. Skip the media changes on Saturday and Sunday.

NOTE: Adjust the split ratio to achieve 70-80% confluence at the time of passage, typically it will be lower than that of daily feed (e.g., if the daily feed is 1:20 every four days, then WF will be ~1:15).

Adaptation from legacy media

For the complete protocol on how to adapt cells cultured in legacy media to BMEM or BMEM Max, visit this page.

Product Use

For Research Use Only. 

Not for diagnostic procedures. 

Reference 
  1. Lyra-Leite, D.M., Copley, R.R., Freeman, P.P., Pongpamorn, P., Shah, D., McKenna, D.E., Lenny, B., Pinheiro, E.A., Weddle, C.J., Gharib, M., et al. (2023). Nutritional requirements of human induced pluripotent stem cells. Stem Cell Reports 18, 1371-1387. https://doi.org/10.1016/j.stemcr.2023.05.004.