IMDM Media Formulation

A highly enriched medium that is formed by the modification in the Dulbecco’s medium [1].

1. Introduction:

Iscove’s Modified Dulbecco’s Medium was first prepared by N.N. Iscove in 1978. He cultured B lymphocytes (stem cells developed from bone marrow) of mice that developed to its maturation stage without even adding serums.
In IMDM media formation, Dulbecco’s Modified Eagle Medium quality was enhanced by adding selenium and in place of serum, it contains albumin, soya bean lipids, and transferrin.
Due to the presence of replacement supplements and selenium, it becomes highly enriched in amino acids, vitamins, and glucose [1].
It is obtainable in the market in different sizes like in the packaging of 250ml, 500ml, 1,000 ml, 10 x 500 ml, and 1 Liter. It is typically packed in sustainable plastic bottles tightly sealed [2].

2. Procedure:

The production procedure of Iscove’s Modified Dulbecco medium with 25mM HEPES buffer and Sodium bicarbonate, Without L-Glutamine (1X Liquid Cell Culture Medium) in the laboratory typically includes a number of essential components which are added in measured quantities. The names and quantities of fundamental components that are combined together to form an IMDM medium are described below in detail [6] [7] [8].

Ingredients	Concentration (mg/L)
Other Components:
D-Glucose	4500
Sodium pyruvate	110
HEPES	5958
Phenol red sodium salt	15.00
Vitamins:
Vitamin B12	0.013
Choline chloride	4.00
D-Biotin	0.013
i-Inositol	7.20
Niacinamide	4.00
D-Calcium pantothenate	4.00
Riboflavin	0.40
Folic Acid	4.00
Thiamine HCl	4.00
Pyridoxal HCl	4.00
Inorganic Salts:
Potassium Chloride (KCl)	330
Sodium Bicarbonate (NaHCO3)	3024
Potassium Nitrate (KNO3)	0.076
Sodium Phosphate (NaH2PO4-H2O)	125
Sodium Chloride (NaCl)	4505
Sodium Selenite (Na2SeO3-5H20)	0.0173
Magnesium sulphate anhydrous	97.7
Sodium dihydrogen phosphate anhydrous	109
Calcium chloride dihydrate	219
Amina Acids:
Glycine	30
L-Alanine	25
L-Valine	93.6
L-Arginine hydrochloride	84
L-Tyrosine disodium salt	104.2
L-Asparagine	25
L-Aspartic acid	30
L-Cystine dihydrochloride	91.24
L-Glutamic acid	75
L-Lysine hydrochloride	146.2
L-Histidine hydrochloride monohydrate	42
L-Isoleucine	104.8
L-Methionine	30
L-Leucine	104.8
L-Phenylalanine	66
L-Proline	40
L-Threonine	95.2
L-Serine	42
L-Tryptophan	16

The core components included in Iscove’s Modified Dulbecco’s medium with 25mM HEPES buffer and Sodium bicarbonate, are the indicator (phenol red), glucose, and without L-Glutamine.
Further 20ml of L-glutamine is added for 1 liter of the medium if required.

3. Specifications:

The specifications of the Iscove’s Modified Dulbecco’s Medium are described below [2] [9] [10].

PROPERTIES	IMDM MEDIA
Physical state	Liquid
Color	Orangish red
Odor	N/A
pH	7.00 -7.60
Impurities	Endotoxins present
Concentration of endotoxins	5EU/ml
Osmolality	280.00 -320.00 mOsm/Kg
Density	N/A
Storage condition	2 to 8 degrees Celsius
Concentration of glucose	High glucose
Buffer	HEPES
Indicator	Phenol red
Glutamine	L-Glutamine
Sodium pyruvate additive	Sodium pyruvate
Sterility	Sterile filtered
Shelf life	12 to 18 months
Shipping condition	Room temperature
Green Features	Sustainable packages
Oxidizing property	None
Chemical stability	Stable

4. Applications:

The practical applications of Iscove’s Modified Dulbecco’s Medium (IMDM Media) are described below [1] [2] [5] [6] [11].

Development of Mammalian Cells:
IMDM helps in the development and maturation of many mammalian tissues or cells like hematopoietic progenitors, Myeloid Cell, and bone marrow cells.
Development of B and T Lymphocytes:
The nutritional supplements present in the Iscove’s Modified Dulbecco’s Medium promote growth and development of B Lymphocytes that derived from bone marrow) and T Lymphocytes that are derived from the thymus.
Precursor Cells of Erythrocytes and Macrophages:
This medium provides essential nutrients for the normal growth of precursor cells of RBC’s (erythrocytes) and macrophages.
Adjustment of pH:
Iscove’s Modified Dulbecco’s Medium adjust pH by using sodium bicarbonate buffer and it also uses Carbon dioxide environment for maintaining the pH.
Cell growth and viability:
Iscove’s Modified Dulbecco’s Medium supports cell growth and viability.
Growth of primary hematopoietic cells: Iscove’s Modified Dulbecco’s Medium helps in the serum-free development of primary hematopoietic cells.
Cell Cultures:
Iscove’s Modified Dulbecco’s Medium rapidly proliferates high-density cell cultures in a 5% carbon dioxide atmosphere.

5. Hazards and Safety Measures:

The hazards and safety measures of Iscove’s Modified Dulbecco’s Medium are described below in detail [3] [4].

Non-Corrosive in nature:
Iscove’s Modified Dulbecco’s Medium is potentially not corrosive in nature.
Contact with eyes:
Iscove’s Modified Dulbecco’s Medium solution is not dangerous for the eyes. Long-term exposure to the eyes may cause eye irritation. Try to protect unexposed eyes by using side shields or goggles. It can cause redness.
- First Aid Measure: If eyes get in contact with the liquid form of Iscove’s Modified Dulbecco’s Medium, wash with plenty of water for 5 to 10 minutes. Open upper and lower eyelids and wash them gently with splashes of water. There is no need of consulting medical advice immediately.
Contact with skin:
Iscove’s Modified Dulbecco’s Medium is not corrosive for the skin. If it gets in contact with the skin, it may cause little or no itching on the skin.
- First Aid Measure: Remove the contaminated clothes and wash them. Wash the exposed area thoroughly from water. Use neutral soaps or liquid soaps for washing the body parts.
Inhalation:
It may not cause irritation and burning sensation in the respiratory system on inhalation of mist, vapors, or dust of Iscove’s Modified Dulbecco’s Medium.
- First Aid Measure: It is unhazardous on inhalation. Properly consult medical advice if ordinarily required.
Ingestion:
If it is ingested mistakenly, it may not cause mild irritation or functional disturbance in the sensitive stomach.
- First Aid Measure: Do not induce vomiting. Immediately wash the mouth of the victim with excess water and enable him to drink two glasses of water. Properly consult medical advice if typically needed.
Safety measures:
Typically wear safety goggles, chemically resistant gloves, and protective clothing. Properly use well-ventilated areas for continuous experimentation and research purposes only.

6. Key Recommendations:

Don’t utilize the specific product if it is expired. Always see the expiry date before handling and using the valuable product.
Properly take care of the storage conditions of the Iscove’s Modified Dulbecco’s Medium.

Animal Cell & Plant Tissue Culture Reagants

IMDM Media Formulation