Culture Media Components & Role in Growth.pptx

Culture Media Components including microbial, mamalian and Plant Tissue Culture with their role in Grwoth

1. Culture Media Components and Their Role in Growth Presented To Dr. Sarfraz Kiani Presented By: Muhammad Sheeraz Majeed SU91-BSMBM-F22-003 BS Microbiology

2. Introduction The culture medium is the single most important factor determining the success of in vitro growth. Culture Medium: Artificial environment providing physical and chemical conditions for growth Types: Microbial, Plant, and Mammalian Culture Media Importance: Used in disease diagnosis, pharmaceutical production, and biological research

3. Microbial Media – Types Based on Physical State Solid: For colonies (agar) Liquid: Broth for suspended growth Semi-solid: Motility testing Based on Composition Defined: Exact composition k nown Complex: Contains extracts (yeast, beef) and e xact composition unknown Based on Function Simple: General purpose Enriched: Extra nutrients for fastidious organisms Selective: Inhibits unwanted microbes Differential: Distinguishes between microbes

4. Fig 1: Different types of Microbial Media

5. Microbial Media – Components & Roles Macronutrints: Carbon sources (sugars, amino acids) → energy Nitrogen sources (peptones, ammonium salts) → proteins & nucleic acids Phosphorus sources (phosphate salts) → nucleic acids & membranes Sulphur sources (amino acids, sulfates) → enzymes & cofactors Micronutrients: Mg²⁺ → enzyme cofactor, membrane stability Ca²⁺ → cell wall formation Fe²⁺/Fe³⁺ → electron transport chain Trace elements (Zn, Mn, Cu) → enzyme activity

6. Other Microbial Media Components Water: Solvent for all biochemical reactions. pH Buffers: Phosphates maintain optimum pH. Vitamins: B-complex (coenzymes in metabolism and redox reactions). Gelling Agent: Agar (from red algae) forms solid surface for colonies. Agarose → neutral polysaccharide Agaropectin → contains sulfate and acidic groups

7. Plant Tissue Culture Media – Types Murashige & Skoog (MS): balanced nutrients and hormones Gamborg’s B5: rich in vitamins and amino acids White’s Medium: root culture (low nutrients) Nitsch’s Medium: anther culture specific hormone balance Woody Plant Medium (WPM): for woody species Fig 2: Plant Tissue Culturing

8. Media Components of Plant Tissue Culture Fig 3: Media Components of Plant Tissue Culture

9. Media Components of Plant Tissue Culture Macronutrients: Nitrogen ( 25–60 mM) → Protein & nucleic acid synthesis Potassium(20–30mM) → Enzyme activation & osmotic regulation P, Ca, Mg ( 1–3 mM) → Energy transfer, cell wall synthesis, cofactor function Micronutrients: Cobalt, Iodine(0.1 μ M )→ Trace enzymatic functions Iron, Molybdenum (1 μ M) → Chlorophyll & electron transport Zinc(5–30 μ M) → Enzyme activation Boron (25–100 μ M) → Cell wall formation Manganese(20–90 μ M) → Photosynthesis & enzyme cofactor

10. Plant Media Components – Vitamins & Hormones Vitamins: Thiamine (B1), Nicotinic acid, Pyridoxine (B6) → coenzymes Amino Acids: Glycine, Glutamine, Casein hydrolysate → nitrogen source Plant Growth Regulators: Auxins: Root initiation, callus formation Cytokinins: Shoot formation, bud proliferation High Auxin: Cytokinin → Rooting media Low Auxin: Cytokinin → Shoot multiplication

11. Fig 4: Auxin: Cytokinin Ratio

12. Plant Media Components – Support & Additives Myo -inositol: Stimulates cell division & membrane synthesis Solidifying Agents: Agar (0.8–1%) preferred Organic Extracts: Coconut milk, yeast extract – growth promotion Antibiotics/PPM: Prevent contamination e.g ; Kanamycin can prevent contamination but may inhibit plant cell growth

13. Mammalian Cell Culture – Media Types Dulbeeco modified Eagle Media ( DMEM ) : High-glucose medium for HeLa & HEK293 cells RPMI 1640: Used for lymphocyte cultures Eagle minimal essential media EMEM: Basic nutrient medium for mammalian cells

14. Media Components of Mammalian Cell Culture Fig 5: Media Components of Mammalian Cell Culture

15. Mammalian Media – Components & Roles Basal Medium: Glucose → main energy source Amino acids → protein synthesis Vitamins → enzyme cofactors Inorganic salts → osmotic balance Serum (FBS): Growth factors (IGF, FGF, EGF) → cell proliferation Hormones → metabolic regulation Lipids → membrane integrity Attachment factors (fibronectin) → cell adhesion Supplements: Antibiotics/antimycotics → contamination control CO₂/Bicarbonate buffer → maintains pH 7.4

16. Functional Roles in Growth Microbial Media: Simple energy sources for rapid growth Plant Media: Hormone-directed development and differentiation Mammalian Media: Mimics internal body fluids; serum-dependent

17. Conclusion Culture media are specialized environments tailored to specific cell types Each component has a precise role in supporting life processes Essential for biotechnology, medicine, and research

18. Refrences 1. Saad & Elshahed (2012) – Plant tissue culture media 2. Waters (2022) – Cell culture medium components 3. Freshney (2015) – Animal cell culture techniques 4. Madigan et al. (2021) – Brock Biology of Microorganisms 5. MicrobiologyInfo.com – Culture media uses

19. Thank You