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ARTIFICIAL CELLS

ARTIFICIAL CELLS. Life is endowed with a mysterious and divine life-force. Jayanti Tokas 1 , Rubina Begum 1 , Shalini Jain 2 and Hariom Yadav 2 1 Department of Biotechnology, JMIT, Radaur 2 NIDDK, National Institute of Health, Bethesda,MD20892, USA Email: yadavhariom@gmail.com.

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ARTIFICIAL CELLS

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  1. ARTIFICIAL CELLS Life is endowed with a mysterious and divine life-force Jayanti Tokas1, Rubina Begum1, Shalini Jain2 and Hariom Yadav2 1Department of Biotechnology, JMIT, Radaur 2 NIDDK, National Institute of Health, Bethesda,MD20892, USA Email: yadavhariom@gmail.com

  2. Artificial Cells • Artificial microscopic structures • Same size as biological cells • Have some of the functional properties of biological cells. • They contain biologically active materials.

  3. Properties of an ideal minimal cell • An information carrying polymer • Transport across the membrane • An external source of chemical energy • A catalytic activity • Growth and Division • Regulation

  4. Hypothetical cell like structure

  5. Membranes • Lipid bilayer vesicles(liposomes)- Multilamellar vesicles(MLVs) • Small unilamellar vesicles(SUVs)-5nm-100nm. • Large unilamellar vesicles(LUVs)-100nm-1µm. Smallest biological cell - 0.2-0.5 µm

  6. Transport across cell membranes • An assisted mechanism is necessary • Carrier molecules • Channels • Pumps

  7. Channels • Neutral peptides • Cyclic peptides • α-hemolysin • A tetrameric channel - M2 protein

  8. Energy Supply • ATP molecules from the environment • Mimic the energy transduction process used by all living cells

  9. Encapsulating Macromolecules • Catalytic activity of the macromolecule is not damaged • Dehydration- re hydration method • Freeze- thaw technique • Injection of molecules

  10. Micro encapsulation • To separate functional macromolecules • Regulate exchange of materials • Support metabolism • Transduce environmental energy into chemical energy • Synthesize a desired biosynthetic product

  11. Challenges for cell encapsulation • Optimize • Biocompatibility • Mass transfer • Stability • Reproducibility • Structural • Functional relationship • Increases Long term stability • Repoducible results

  12. A number of commercial machines are available for automatic production of artificial cells

  13. Preparation of Artificial cell

  14. Two step Method

  15. Contd..

  16. Contd...

  17. Artificial cells • Like biological cells, Artificial Cells function with content retained inside to – • Act on outside permanent molecules • Release products of interaction

  18. Dimensions of Polymeric Artificial Cells • Macro dimensions • For genetically engineered cells, stem cells, other cells, tissues, microorganisms, etc. • Micron dimensions • For enzymes, genetically engineered microorganisms and other microorganisms, peptides, etc. • Nano dimensions • For blood substitutes, enzymes, peptides, magnetic materials, drugs, etc. • Molecular dimensions • For blood substitutes, crosslinked enzymes, conjugated proteins, etc.

  19. APPLICATIONS

  20. Artificial cells containing biological cells • Microencapsulated islets for Diabetes Mellitus • Microencapsulated hepatocytes for liver failure *Stem cells

  21. Artificial cells containing genetically engineered cells • Beta-endorphin secreting cells • Recombinant ciliary neurotrophic factor secreting cells • Oral therapy for Uremia- E.coli DH 5 cells

  22. Artificial cells containing enzymes • Treatment of enzyme deficiency diseases * Phenylketonuria * Lesch Nyan Disease * Skin Cancer Melanoma • Urea removal

  23. Artificial cells containing microorganisms • Microencapsulation of cholesterol removing microorganisms- Pseudomonas pictorum

  24. Artificial Cells With Nano-Dimensions

  25. Typical nano-dimension artificial cells of 80nm mean diameter

  26. Artificial RBC(Hb +Enz) with all Biochemical properties

  27. Artificial cells in Molecular Dimensions • As Oxygen Carrier

  28. Contd..

  29. Aid in Cancer Therapy

  30. Polymeric Artificial cells Containing Cells

  31. Contd..

  32. Contd..

  33. Bioactive sorbants • Activated charcoal and resins • Ultra thin coating of colloidal membrane • Detoxify the blood – hemoperfusion. • McGill Artificial Cells and Organ Research Centre in the late 1960s.

  34. Hemoperfusion Patient’s blood Charcoal-filled artificial cells Toxins in the blood to enter the cells Adsorbed by the charcoal.

  35. Artificial cells as drug delivery vehicles Drug (biodegradable membrane) fusion Target tissue membrane degraded Drug released

  36. Artificial cells as biosensors Artificial cells analytes • signal Coenzyme-depleted enzyme - glucose oxidase

  37. Artificial Cell Membrane Holds Promise For Medical Use • Detecting Flue Virus • Anti-inflammatory Agent

  38. Tailored Glycolipids(sialic acid) Bind Specific Protein (Coat proteins of Influenza virus) Pink film

  39. Inflammatory Agents Infection/Injury Blood Vessels Produce Receptors WBC/Neutrophils Bind

  40. Contd.. Excess * Rheumatoid Arthritis * Septic Shock * Clamping of Blood Vessels During Surgery

  41. Polymerized Membrane Sugars Prevents attachment to Blood Vessels

  42. Artificial red blood cells • Modified hemoglobin * high oxygen carrying capacity * do not have blood group antigens * longer half life * non toxic

  43. E-Pure Water Extracting Hb from RBC’s • RBC’s contain Hb which transport O2 through body • RBC’s are lysed with E-Pure water to extract Hb

  44. Modified Hemoglobin • I generation • II generation • III generation

  45. + Cross-linker In body Why Cross-link Hb? • Hemoglobin must be cross-linked when placed in the blood stream. • Hb breaks into dimers which can travel through capillary pores (holes) and cause death. Cross-linked Hb can no longer travel through pores Hb dimers are small enough to travel through pores

  46. Present status • Hemoperfusion for acute poisoning (clinical trials) • Hemoperfusion for aluminium and iron overload (clinical trials) • Red blood cell substitute (clinical trials) • Diabetes mellitus animal experiments (clinical trials) • Drug delivery systems • Artificial liver support (experimental) • Hereditary enzyme deficiency (clinical trials)

  47. LATEST BREAKTHROUGH • NASA supported Researches for dehydrated blood Supplies • Artificial Sperm • Artificial Human Eggs Possible In 5 years

  48. Artificial Human Egg • Haploidisation • No Cloning • Production of Reconstituted Egg

  49. Procedure Woman’s Somatic Cell *Remove Nucleus *Transfer Shelled Out Oocyte * Problem

  50. Artificial Sperm * Dr. Orly Lacham Kalpan succeeded in fertilizing a normal egg with an artificial sperm. * Embryo Developed normally in Lab

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