OPTI-MEM®

 I Reduced Serum Medium modification of MEM (Eagle's)

CAUTION: Human origin materials are non-reactive (donor level) for anti-

HIV 1 & 2, anti-HCV and HBSAg.  Handle in accordance with

established bio-safety practices.

Cat. No.: 31985                       100 mL  500 mL

Component-Deficient Media:

Cat. No.: *11058            500 mL without phenol red

          51985                500 mL  with GLUTAMAXI

Storage Condition: 2 to 8°C, in the dark.

Shelf Life: See product label for expiration date.

NOTE: For catalog number 11058 please also refer to Component-Deficient

Medium Supplementation below.

OPTI-MEM I is a versatile, chemically-defined medium, formulated to reduce significantly the

amount of serum required for cultivating mammalian cells in vitro.

OPTI-MEM I is a multi-purpose medium that has proven useful in reducing serum requirements for a wide variety of cell lines and applications.  It has been shown effective in the growth and

maintenance of both adherent and non-adherent cell lines.  When supplemented with 2-4% fetal bovine serum or alternative sera, OPTI-MEM I supports proliferative rates and maximal cell

densities comparable to, and in some cases superior to, conventional media supplemented with 10% fetal bovine serum.  Relatively non-fastidious cell lines may be maintained in long-term culture with even more substantial serum reduction.  If using adherent cell lines and less than 2% serum

supplementation or in an agitated system such as in roller bottles, the medium should be further

supplemented with 100 mg/L CaCl2.

The versatility of OPTI-MEM I in the propagation of various cell types makes this medium the optimal choice for many cell culture requirements.

The raw materials used to produce OPTI-MEM I are screened by GIBCOTM rigorous quality standards.  The complete medium is performance tested to ensure the lot-to-lot consistency

required for research and production applications.

The shelf life of OPTI-MEM I is comparable to conventional glutamine-containing formulations.  This stability permits laboratories to stock and maintain inventories of OPTI-MEM I compatible with routine laboratory practices.

Formulation

OPTI-MEM I is a modification of Eagle's Minimal Essential Medium, buffered with HEPES and sodium bicarbonate, and supplemented with hypoxanthine, thymidine, sodium pyruvate, L-glutamine or GLUTAMAX, trace elements and growth factors.  The protein level is minimal (15 µg/mL), with insulin and transferrin being the only protein supplements.  Phenol red is included at a reduced concentration as a pH indicator.  OPTI-MEM I may be supplemented with 2-mercaptoethanol prior to

use.  CaCl2 level is reduced to 99.9 mg/L for convenience in working with hybridomas.

Instructions for Use

                  Conversion to OPTI-MEM I

Converting to OPTI-MEM I is easy.  For most applications, no weaning procedures are necessary to

attain 50% reduction in serum supplementation when converting to OPTI-MEM I.  The conversion can be made by simply centrifuging the cells, decanting the supernatant, and resuspending them in

OPTI-MEM I with the reduced serum supplementation.  (See the tables in the Applications section for typical serum lementation reduction).  Additional serum reduction may be realized with

minimal weaning.  The optimal serum supplementation for each specific application should be determined based on the performance characteristics expected (growth promotion, secondary

metabolite production, etc.).  Extended use of OPTI-MEM I in the maintenance of cell lines has shown no loss of viability or growth rate.  Keep OPTI-MEM I away from direct light.

            Use of 2-Mercaptoethanol

2-mercaptoethanol is a reducing agent which has been shown to enhance growth-1 , plating efficiency-2  and/or antibody synthesis-3  of specific cell lines.  The kinetics of this phenomenon are not fully understood.  Most applications do not require supplementation with 2-mercaptoethanol.

Since OPTI-MEM I is functional in a broad range of applications with numerous cell types, it is prudent to uate the utility of 2-mercaptoethanol supplementation based on the specific

application.

2-mercaptoethanol is available separately as a 1000X concentrate (5.5x10 -2M) in Dulbecco's Phosphate Buffered Saline without calcium and magnesium 21985, 50 mL.  It may be added to

OPTI-MEM I immediately prior to use.  OPTI-MEM I is stable for two months following 2-mercaptoethanol addition, provided that this period does not exceed the expiration date stated on

the OPTI-MEM I or 2-mercaptoethanol labels.

If desired, aseptically add 0.5 mL 2-mercaptoethanol (1000X) to each 500 mL of OPTI-MEM I.  Do not pipette by mouth.  Mix thoroughly by swirling (do not shake).  Store unused portions of 2-mercaptoethanol tightly capped in the original container.  The final concentration of 2-mercaptoethanol in OPTI-MEM I will be 5.5x10-5M.