1. Jayanti Tokas1, Rubina Begum1, Shalini Jain2 and Hariom Yadav2
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

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 - 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
Cell Content
Aim
Cells and Tissues
Pancreatic cells
Feedback controlled secretion of insulin for diabetes mellitus
Hepatocytes
To support liver function in liver failure
Kidney cells
To secrete erythropoietin to treat anemia
Parathyroid Cells
To secrete parathyroid hormone to treat hypoparathyroidism

31. Genetically engineered cells
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 hepatocytes
Increase duration of viability and function of hepatocytes

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

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
analytes
signal
Coenzyme-depleted enzyme - glucose oxidase

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

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

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

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

41. Prevents attachment to
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. 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
E-Pure Water

44. Modified Hemoglobin
I generation
II generation
III generation

45. 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-linker
In body
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 Artificial Sperm
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 * Problem
*Remove Nucleus
*Transfer
Shelled Out Oocyte
* Problem

50. * Embryo Developed normally in Lab
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.

52. THANK YOU