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ARTIFICIAL CELLS Jayanti Tokas 1, Rubina Begum 1, Shalini Jain 2 and Hariom Yadav 2 1 Department of Biotechnology, JMIT, Radaur 2 NIDDK, National Institute.

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Presentation on theme: "ARTIFICIAL CELLS Jayanti Tokas 1, Rubina Begum 1, Shalini Jain 2 and Hariom Yadav 2 1 Department of Biotechnology, JMIT, Radaur 2 NIDDK, National Institute."— Presentation transcript:

1 ARTIFICIAL CELLS 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 Life is endowed with a mysterious and divine life-force

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 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 - M 2 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  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. Dimensions of Polymeric Artificial Cells

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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 Cell ContentAim Cells and Tissues Pancreatic cellsFeedback controlled secretion of insulin for diabetes mellitus HepatocytesTo support liver function in liver failure Kidney cellsTo secrete erythropoietin to treat anemia Parathyroid CellsTo secrete parathyroid hormone to treat hypoparathyroidism

31 Contd.. Genetically engineered cells Mouse myoblast (mGH gene) SK2 hybridoma cells Secrete mGH for dwarfism Secrete anti-hIL-6 monoclonal Abs for IgG1 plasmacytosis Mouse myoblasts (human factor IX geen) Secrete human factor IX for hemophilia B Hamster kidney cells (CNTF gene)Secrete ciliary neutrotrophic factor for amytrophic lateral sclerosis Neuro2A cells (pro-piomelaocortin gene) Secrete β-endorphin for pain treatment Stem cells Stem cells plus hepatocytesIncrease duration of viability and function of hepatocytes

32 Contd.. Microorganisms Pseudomonas pictorum To remove Erwinia herbicolaTo convert ammonia pyruvate and phenol into lysine and DOPA CDH5 E.coliUrea removal Metabolic induction of Lactobacilus delbruecki Urea removal

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) Specific Protein (Coat proteins of Influenza virus) Pink film Bind

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

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

41 Polymerized Membrane Prevents attachment to Blood Vessels Sugars

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

43 Extracting Hb from RBC’s  RBC’s contain Hb which transport O 2 through body  RBC’s are lysed with E-Pure water to extract Hb E-Pure Water

44 Modified Hemoglobin  I generation  II generation  III generation

45 Why Cross-link Hb? + Cross-linker In body  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. Hb dimers are small enough to travel through pores Cross-linked Hb can no longer 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  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 Shelled Out Oocyte *Remove Nucleus *Transfer * Problem

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

51 Conclusion Artificial cells allow to combine properties of biological systems for therapeutic and diagnostic applications. Artificial cell is an attempt to mimic some of the biological processes of a real cell, it may ultimately prove most useful as a partial substitute for animal cells.

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