Solid organ fabrication is an ultimate goal of Regenerative Medicine. Since the introduction of Tissue Engineering in 1993, significant advancements have been made to regenerate in vitro culture or tissue platforms. Relatively simple flat or tubular organs are already in (pre)clinical trials . There are two emerging technologies for solid organ fabrication. One is decellularization of cadaveric organs followed by repopulation with terminally differentiated or progenitor cells. The other is 3D bioprinting to deposit cell-laden bio-inks to attain complex tissue architecture.

1. Chasing The Holy Grail Of
Regenerative Medicine

2. Content
• Regenerative medicine
• Areas of Regenerative Medicine
• Introduction
• Extra Cellular Matrix
• Objectives
• Decellularization
• Approaches in the field
• Accomplished To Date
• Pro & cons
• Hurdles
• Conclusion
• Bibliography

3. Characterizing
• Regenerative Medicine : Regenerative medicine is a broad
definition for innovative medical therapies that will enable the
body to repair, replace, restore and regenerate damaged or
diseased cells, tissues and organs.
2. Tools and Procedures :
• Tissue Engineering: Tissue Repair/Replacement and Lab
Grown Organs
• Technologies
a. Stem cells
b. Natural and Synthetic Scaffolds
c. 3-D Printing and Chip Technologies

4. Areas of Regenerative Medicine
1. Artificial Organs: Medical Devices (Lab Grown
Bladder)
2.Tissue Engineering & Biomaterials Scaffolds

5. Areas of Regenerative Medicine
3. Cellular Therapies
• Use of Stem Cells (From Patient)
• Development of Regenerative Medicine Treatments.
• Enhance Regeneration of Tissues and Organs.
4. Clinical Trials
• Many Currently in Progress.
• NIH and Private Organizations.

6. Introduction
• Solid organ fabrication is an ultimate goal of
Regenerative Medicine. Since the introduction of
Tissue Engineering in 1993, significant advancements
have been made to regenerate in vitro culture or
tissue platforms. Relatively simple flat or tubular
organs are already in (pre)clinical trials .
• There are two emerging technologies for solid organ
fabrication.
1. One is decellularization of cadaveric organs
followed by repopulation with terminally
differentiated or progenitor cells.
2. The other is 3D bioprinting to deposit cell-laden
bio-inks to attain complex tissue architecture.

7. People Die Every Year
Sales
Coronary heart failure
Chronic lung diseases
Kidney failure
Liver diseases

8. Treatments For Organ Failure
• 1) Frequent Treatment
Allogeneic organ transplantation
Xenogeneic organ transplantation
Artificial organs
• Disadvantages
1. Scarcity of donors
2. Body immune response
3. Thrombogenicity

9. Tissue Engg Approach
Bioartificial organs
• Patients own cell
• Organ development
• Transplantation
• Scaffold (ECM)

10. Tissue Engg Approach

11. Extra Cellular Matrix
• Framework for cell
binding and organ
formation
• Made of
proteins,glycoproteins ,
proteoglycans
• Signals for cell growth&
proliferate
• Supports cell adhesion
migrations differation and
proliferate.

12. Objectives
1. Decellularization
2. Obtaining the appropriate type and no of cells.
3. Reseeding the ECM with cells
4. Organ maturation
5. Transplantation & in vivo testing

13. Decellularization
• It began with great promise to regenerate cadaveric
organs while overcoming transplant rejection and
possibly alleviating perpetual shortage of donated
organs.
• It can be done with organs harvested from pigs or
newly dead organs.

14. • Cells in these organs are stripped away through
decellularzation process.
• Scientist wash the cell away with the help of a special
detergent and chemicals to wash away its DNA, lipids
,soluble proteins ,sugar and other cellular material .
• After washing we are left with scaffold of original
tissue .

15. • The sterile scaffold is then seeded with stem cells
taken from patient ie progenitor cells
• The stem cells are preprogrammed to become
specialized cell depending upon the organ.
• All this process helps the patient to avoid the risk if
rejection by the recipient immune response.

17. Decellularization Of Tissues And
Organs
Physical Enzymatic Chemical
Mechanical agitation Trypsin Alkaline/acid
Freeze/thaw Endonucleases Hypotonic and
hypertonic solutions
Sonication Exonucleases EDTA, EGTA
Nonionic detergents
Triton X-100
Ionic detergents
Sodium dodecyl sulfate (SDS) Triton X-200
Zwitterionic detergents
CHAPS Sulfobetaine-10 and -16 (SB-10, SB-16)
Tri(n-butyl)phosphate
Abbreviations: CHAPS, 3-[(3-
cholamidopropyl)dimethylammonio]-1-
propanesulfonate; EDTA, ethylene
diamine tetraacetic acid; EGTA, ethylene
glycol tetraacetic acid

18. Building Scaffolds
• Utilizing xenogeneic or non-transplantable organs as
scaffolds for re-building tissues with stem cells

19. What it does to build a solid organ
A scaffold
• Biologic
(ecm)
• Synthetic
• Bioprinting
Billions of cells
• Bone marrow
• Blood
• iPS cells
• ES cells2+
Ability to put it
together
• Pump
function
• Metabolism
• physiology

20. Cell Sources
• Embryonic stem cells
• Fetal cells
• Adult derived stem cells / progenitor cells
• Umbilical cord blood cells
• Organ derived stem cells
• Organ derived progenitor cells
• iPS

21. Approaches in the field
• Haralad C ott et. al .(2008)perfusion decelluarized
matrix using natures platform to engineer a
Bioartificial heart ie rat heart.
Perfusion decellularization with
different detergents
1% PEG
After 12 hrs
1% Triton X-100
After 12 hrs
1% SDS
After 12 hrs best result

22. Approaches in the field
• Haralad C ott et. al .(2008)perfusion decelluarized
matrix using natures platform to engineer a
bioartificial heart ie rat heart.
Recellularization with endothelial
cells after 7 days
• 550.7 ± 99.0 endothelial
cells/mm2 on the endocardial
surface
• 264.8 ± 49.2 endothelial
cells/mm2 within the vascular tree
Recellularization with neonatal
cardiomyoctes
• After 8 days the heart s showed
contructs and electric responses (
2% of adult rat heart function

23. Approaches in the field
• Jeremy J Song et al(2013) Regeneration &
experimental orthotopic transplantation of a
bioengineered kidney.
Perfusion decellularization with SDS
detergent for 12 hrs
Reseeding with 50 ×106 epithelial cells
Orthotopic transplantation and urine
production

24. Accomplished To Date
Whole Heart Decellularization
• Designing a bioreactor
• Minimizing the total decellularization time to
time to 1 day and SDS contact time for 4 hrs
procine hearts.

25. Accomplished To Date
• Arota Decllularization / Recellularization Bioreactor
• Porcine Aortic Endothelial Cell Culture

26. Accomplished To Date
• Thrombosis Assay
3 hr - Static Blood Thrombosis Assay

27. Creating a Beating Heart in the
Lab
• Decellularization Recellularization

28. Creating a Beating Heart in the Lab
Step 1. Cardiac Patches as a tool for repair or delivery
(porcine)

29. • Step 2. Revascularized hearts
• Step 3. Bio-Artificial Hearts for Transplant

30. Pro’s & Con’s
• Pro’s
1. Most nature simulating scaffolds in terms of
composition & mechanical properties
• Con’s
1. Inhomogeneous distribution of cells
2. Difficulty in retaining all ECM
3. Immunogenicity upon incomplete Decellularization
Preferred application : tissue with high ECM content
load bearing tissue.

31. Hurdles
• Organ complexity Atria, Ventricle, Pacemaker,
Neurons, Fibroblasts, Stem cells
• Billions of cells at an affordable cost
• Once we have cells … organ potency
• Longevity – years and years
• Endogenous responsively/repair
• Physiologic response Progenitor cells Cardiac stem
cells Endothelial cardiomyocytes Smooth muscle cells
Numbers Types

32. Conclusions
• The unmet need is organs for transplant
• dECM provides optimal advantages as a scaffold
• The door is open for complex human organ
engineering
• Regenerative medicine is coming of age
• The rate- limiting step is CELLS

33. References
• Decellularized Tissue Engineering
• Repopulation of decellularized whole organ scaffold
using stem cells: an emerging technology for the
development of neo-organ
• Perfusion decellularization of whole organs.
• Bioartificial Heart: A Human-Sized Porcine Model –
The Way Ahead
• Perfusion-decellularized matrix: using nature's
platform to engineer a bioartificial heart
Harald C Ott1, Thomas S Matthiesen2, Saik-Kia Goh2,
Lauren D Black3, Stefan M Kren2, Theoden I Netoff3 &
Doris A Taylor2,4

34. References
• https://www.researchgate.net/profile/Korkut_Uy
gun/publication/50591416_Whole-
Organ_Tissue_Engineering_Decellularization_and
_Recellularization_of_Three-
Dimensional_Matrix_Scaffolds/links/0c9605373b
8099c6af000000.pdf
• https://link.springer.com/chapter/10.1007/978-
981-10-3328-5_5
• https://www.hindawi.com/journals/bmri/2017/9
831534/