Human Tubal Fluid (HTF) Media & Modifi ed Human Tubal Fluid (mhtf) Medium with Gentamicin

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1 Human Tubal Fluid (HTF) Media & Modifi ed Human Tubal Fluid (mhtf) Medium with Gentamicin HTF Media are intended for use in assisted reproductive procedures which include gamete and embryo manipulation through Day 3 of development. HTF Medium is a synthetic, defined solution for use as a culture media through Day 3 of human embryo development as well as the processing of gametes. HTF is bicarbonate-based and is designed for use in a CO 2 HTF Medium, Catalog #90125 Available as liquid medium in 100 ml configuration. Custom configurations are available Has a shelf life of 64 days from the date of manufacture Complete HTF Medium with SSS, Catalog #9922 * Available as liquid medium in 2 x 20 ml configuration Ready to use, already supplemented with protein (10% SSS) for a final total protein concentration of 6 mg/ml mhtf Medium is a synthetic, defined solution for procedures which include the retrieval, handling and transfer of human gametes and embryos. mhtf uses a buffering system composed of a 21mM HEPES and 4 mm Sodium Bicarbonate combination. This buffering system does not require the use of a CO 2 mhtf Medium, Catalog #90126 Available as liquid medium in 100 ml configuration. Custom configurations are available Has a shelf life of 180 days from the date of manufacture HTF Sodium Chloride 97.8 Potassium Phosphate, Monobasic 0.37 Sodium Bicarbonate 25.0 Glucose 2.7 Protein Supplementation None (User Option) Complete HTF with SSS Component mm Sodium Chloride Potassium Chloride 4.22 Magnesium Sulfate, Anhydrous 0.18 Potassium Phosphate 0.33 Calcium Chloride 1.84 Sodium Bicarbonate Glucose 2.50 Sodium Pyruvate 0.30 Sodium Lactate Phenol Red 4.5 mg/l Gentamicin 9 μg/ml Human Serum Albumin 5 mg/ml Globulins 1 mg/ml Modified HTF Component mm Sodium Chloride 97.8 Potassium Phosphate, Monobasic 0.37 Sodium Bicarbonate 4.0 HEPES 21.0 Glucose 2.78 Protein Supplementation None (User Option) Each lot of HTF media receives a s CATALOG #9922, & Rev.8

2 Single Step Medium Single Step Medium (SSM ) is intended for use as a culture medium for human gametes and embryos during fertilization and growth of embryos to Day 5/6 of development. Single Step Medium, Catalog #90161 Shelf life is 90 days from date of manufacture when stored 2-8 C For use in a CO 2 incubator SSM is a chemically defined complex medium containing glucose, alanyl-glutamine (as a stable source of glutamine), optimized concentrations of essential and non-essential amino acids and citric acid as an antioxidant Can be used from fertilization through Day 5/6 of development Only one medium to order, inventory and store Produced with Irvine Scientific s WFI Grade Water. Component mm Sodium Chloride Potassium Chloride Potassium Phosphate Calcium Chloride, Anhydrous Magnesium Sulfate, Anhydrous Sodium Bicarbonate Sodium Pyruvate Glucose Sodium Citrate Sodium Lactate (D/L) EDTA, Disodium, Dihydrate μm Alanyl-glutamine Alanine Arginine Asparagine Aspartic Acid Cysteine Glutamic Acid Glycine Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Proline Serine Taurine Threonine Tryptophan Tyrosine Valine Phenol Red mg/l Gentamicin μg/ml Each lot of Single Step Medium receives a s CATALOG #90161 Rev.2

3 P-1 (Preimplantation Stage One) Media P-1 Medium, the Gold Standard, are intended for use in culturing human gametes and embryos during fertilization and growth through Day 3 of development. P-1 Medium was developed by Dr. Thomas B. Pool, Fertility Center of San Antonio, based upon the original HTF formula but modified to provide a glucose-free and phosphate-free environment. This formulation also contains taurine, an amino acid shown to have protective and embryogenic functions. P-1 Medium is bicarbonate-based and is designed for procedures utilizing a CO 2 P-1 Medium, Catalog #99242 Available as liquid medium in 100 ml configuration. Has a shelf life of 64 days from the date of manufacture Requires Protein Supplementation Complete P-1 Medium with SSS, Catalog #9926 ** Ready to use, supplemented with protein (10% SSS), for a final total protein concentration of 6 mg/ml. Complete P-1 Medium with DSS, Catalog #9910 ** Ready to use, supplemented with protein (10% DSS), for a final total macro molecule concentration of 7 mg/ml. P-1 Medium is a simple salt solution that does not contain glucose.. Formulated with Irvine Scientific s WFI Grade Water. Chemicals used in Irvine Scientific s reproductive products meet USP and ACS standards, where available. P-1 Medium* Sodium Chloride Sodium Bicarbonate 25 Taurine 0.05 Sodium Citrate 0.15 mg/l Complete P-1 Medium with SSS Component mm Sodium Chloride Potassium Chloride Magnesium Sulfate, Anhydrous Calcium Chloride, Anhydrous Sodium Bicarbonate Sodium Pyruvate Sodium Lactate Taurine Sodium Citrate mg/l Phenol Red mg/l Gentamicin μg/ml Human Serum Albumin mg/ml Globulins mg/ml Complete P-1 Medium with DSS Component mm Sodium Chloride Potassium Chloride Magnesium Sulfate, Anhydrous Calcium Chloride, Anhydrous Sodium Bicarbonate Sodium Pyruvate Sodium Lactate Taurine Sodium Citrate mg/l Phenol Red mg/l Gentamicin μg/ml Human Serum Albumin mg/ml Dextran mg/ml *Original formulation provided by Thomas B. Pool, Ph.D., Fertility Center of San Antonio. * Each lot of P-1 media receives a complete laboratory evaluation including mouse embryo testing, endotoxin level, ph, osmolality and sterility testing. All results are provided in a lot-specifi c Certifi cate of Analysis. s CATALOG #99242, 9910 & 9926 Rev.5

4 Early Cleavage Media Early Cleavage Media (ECM ) are intended for use in culturing human gametes during fertilization (IVF) and growth of embryos through Day 3 of development. ECM Medium is a modification of P-1 Medium that contains glucose, alanyl-glutamine and EDTA, components shown to benefit human embryo development 1-6. This medium is an alternative to P-1 Medium for those clinicians who prefer the presence of glucose during fertilization and early cleavage stages of embryo development. ECM is bicarbonate based and is designed for procedures utilizing a CO 2 ECM, Catalog #90138 Has a shelf life of 90 days from date of manufacture Lyophilized ECM Kit, Catalog #90150 Available in 30 ml final configuration Has a shelf life of 1 year from date of manufacture Kit Components: - One (1) Vial of Lyophilized Powder - One (1) Vial of Resuspension Solution Complete ECM with SSS, Catalog #90140 * Supplemented with protein (10% SSS) for a final total protein concentration of 6 mg/ml Complete ECM with DSS, Catalog #90142 * Supplemented with protein (10% DSS) for a final total macromolecule concentration of 7 mg/ml The only culture media system available worldwide as a liquid medium and lyophilized powder Early Cleavage Medium (ECM) is a simple salt solution that includes glucose, alanyl-glutamine and EDTA Formulated with WFI grade water Stored at 2-8 C Liquid Media Formulation: Glucose 0.50 Sodium Chloride Potassium Chloride 2.50 Calcium Chloride, Anhydrous 1.70 Sodium Bicarbonate 25.0 Sodium Lactate (D/L) Alanyl-glutamine 0.50 Taurine 0.05 Sodium Citrate 0.15 mg/l EDTA, disodium, dehydrate 10 μm Phenol Red 4.8 mg/l Resuspension Solution Formulation: SodiumBicarbonate 25.0 Each lot of ECM media receives a s CATALOG #90138, 90150, & Rev.7

5 MultiBlast Media MultiBlast Media are intended for use in culturing human embryos from Day 3 to blastocyst (up to Day 6). Embryos should first be cultured to Day 3 in P-1 Medium or Early Cleavage Medium (ECM ). MultiBlast Medium is formulated to provide a complex array of components required for ongoing development of embryos to the blastocyst stage. This optimized formulation is based upon recent studies and publications. The component concentrations have been selected based upon those present in the physiological environment as well as experimentally optimized values shown to benefit and improve blastocyst development. MultiBlast Medium is bicarbonate based and is designed for use in a CO 2 MultiBlast Medium (liquid), Catalog #90139 Has a shelf life of 90 days from date of manufacture Lyophilized MultiBlast Kit, Catalog # Available in 30 ml final configuration Has a shelf life of 1 year from date of manufacture Kit Components: - One (1) Vial of Lyophilized Powder - One (1) Vial of Resuspension Solution Complete MultiBlast Medium with SSS, Catalog #90141 * Has a shelf life of 120 days from the date of manufacture Already supplemented with protein (10% SSS) for a final total protein concentration of 6 mg/ml Complete MultiBlast Medium with DSS, Catalog #90143 * Has a shelf life of 120 days from the date of manufacture Already supplemented with protein (10% DSS) for a final total macromolecule concentration of 7 mg/ml The only culture media system available worldwide as a liquid medium and lyophilized powder MultiBlast Medium is a chemically defined complex medium containing glucose, alanyl-glutamine, optimized concentrations of essential and non-essential amino acids and citric acid as an antioxidant Formulated with WFI grade water Stored at 2-8 C Liquid Medium Formulation: Component. mm Sodium Chloride Potassium Chloride 2.5 Potassium Phosphate Calcium Chloride, Anhydrous 1.7 Magnesium Sulfate, Anhydrous. 0.2 Sodium Bicarbonate Sodium Pyruvate 0.2 Glucose. 3.0 Sodium Citrate Sodium Lactate (D/L) 20 Alanyl-glutamine 1.0 Alanine Arginine 0.3 Asparagine Aspartic Acid Cysteine Glutamic Acid 0.05 Glycine Histidine 0.1 Isoleucine 0.2 Leucine Lysine 0.2 Methionine 0.05 Phenylalanine. 0.1 Proline 0.05 Serine Taurine Threonine Tryptophan 0.02 Tyrosine. 0.1 Valine 0.2 Phenol Red 4.8 mg/l Gentamicin 10 μg/ml Resuspension Solution Formulation: Component mm Sodium Bicarbonate 25.0 Each lot of MultiBlast Media receives a s CATALOG #90139, 90141, & Rev.7