Maryse de la Giroday 6-week course SFU Liberal Arts & Adults 55+ program

1. Bioelectronics, Medical Imaging and Our BodiesWeek 6: Growing organs (regenerative medicine) and human-on-a-chip Maryse de la Giroday 6-week course SFU Liberal Arts & Adults 55+ program

2. 3D Printing of biological tissue (2011) • http://channel.nationalgeographic.com/channel/explorer/videos/the-human-cell-printer/

3. Cornell University’s bioengineered ear 2013 (1 of 7) • To make the ears, Bonassar and colleagues started with a digitized 3-D image of a human subject’s ear and converted the image into a digitized “solid” ear using a 3-D printer to assemble a mold.

4. Cornell University’s bioengineered ear 2013 (2 of 7) • They injected the mold with collagen derived from rat tails, and then added 250 million cartilage cells from the ears of cows. This Cornell-developed, high-density gel is similar to the consistency of Jell-O when the mold is removed. The collagen served as a scaffold upon which cartilage could grow.

5. Cornell University’s bioengineered ear 2013 (3 of 7) • The process is also fast, Bonassar added: “It takes half a day to design the mold, a day or so to print it, 30 minutes to inject the gel, and we can remove the ear 15 minutes later. We trim the ear and then let it culture for several days in nourishing cell culture media before it is implanted.” [implanted?]

6. Cornell University’s bioengineered ear 2013 (4 of 7) • Bonassar and [Jason] Spector have been collaborating on bioengineered human replacement parts since 2007 • The researchers specifically work on replacement human structures that are primarily made of cartilage — joints, trachea, spine, nose — because cartilage does not need to be vascularized with a blood supply in order to survive.

7. Cornell University’s bioengineered ear 2013 (5 of 7) • They are now looking at ways to expand populations of human ear cartilage cells in the laboratory so that these cells can be used in the mold, instead of cow cartilage. • “Using human cells, specifically those from the same patient, would reduce any possibility of rejection,” Spector said.

8. Cornell University’s bioengineered ear 2013 (6 of 7) • He added that the best time to implant a bioengineered ear on a child would be when they are about 5 or 6 years old. At that age, ears are 80 percent of their adult size. • If all future safety and efficacy tests work out, it might be possible to try the first human implant of a Cornell bioengineered ear in as little as three years, Spector said..

9. Cornell University’s bioengineered ear 2013 (7 of 7) • http://www.frogheart.ca/?p=9310 (Feb. 2013) • Paper: • Reiffel AJ, Kafka C, Hernandez KA, Popa S, Perez JL, et al. (2013) High-Fidelity Tissue Engineering of Patient-Specific Auricles for Reconstruction of Pediatric Microtia and Other Auricular Deformities. PLoS ONE 8(2): e56506. doi:10.1371/journal.pone.0056506 [open access: http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0056506]

10. 2011 vs 2013 bioengineered ears at Cornell • What is the difference?

11. Beyene and the synthetic trachea (1 of 6) • AndemariamTeklesenbetBeyene • Synthetic trachea transplant 2011 • http://www.frogheart.ca/?p=4078 • Student in Iceland, flown to Sweden for transplant by surgeon, Paolo Macchiarini, who has a double appt. Karolinska Institute in Sweden and University College in London, UK

12. Beyene and the synthetic trachea (2 of 6) • Alexander Seifalian (UCL Division of Surgery & Interventional Science; professor of nanotechnology and regenerative medicine at University College London, UK), • created a glass mold of the patient’s trachea from X-ray computed tomography (CT) scans of the patient.

13. Beyene and the synthetic trachea (3 of 6) • In CT, digital geometry processing is employed to generate a 3D image of the inside of an object from a large series of 2D X-ray images taken around one single axis of rotation

14. Beyene and the synthetic trachea (4 of 6) • Seifalian & associates manufactured a full size y-shaped trachea scaffold • Scaffold was built using a novel porous nanocomposite polymer • The composite has millions of little holes(pores),providing a place for the patient’s stem cells to grow roots.

15. Beyene and the synthetic trachea (5 of 6) • The team cut strips of the novel nanocomposite polymer and wrapped them around the glass mold creating to create cartilage rings that confer structural strength to the trachea • Then off to Karolinska where stem cells are harvested and grown in a bioreactor

16. Beyene and synthetic trachea (6 of 6) • During a 12-hour operation Professor Macchiarini removed all of the tumour and the diseased windpipe and replaced it with the tailor-made replica [now covered with tissue grown from the patient's bone marrow tricked into growing like cells found in a trachea

17. Contrast ear story and trachea story • Scan of ear? • CT scan of trachea • ‘Hybrid’ scaffolding (hydrogel and extracellular matrix [cows] and collagen from rat tails) • Nanocomposite scaffolding • Tissue engineering generally requires scaffolding

18. Regenerating a finger and the extracellular matrix (1 of 4) • http://www.youtube.com/watch?v=gwR8rEcVu7c • Stem cells may be needed for regrowth, e.g., stem cells in nail bed may be necessary (at this time) for regenerating fingers (http://www.livescience.com/37380-nail-cells-regenerate-lost-fingers.html)

19. Regenerating a finger and the extracellular matrix (2 of 4) • March 2008 article (Wyatt Anderson) (http://www.cbsnews.com/news/medicines-cutting-edge-re-growing-organs/) • In 2005, Lee Spievack sliced off the tip of his finger in the propeller of a hobby shop airplane

20. Regenerating a finger and the extracellular matrix (3 of 4) • Spievack's brother, Alan, a medical research scientist, sent him a special powder and told him to sprinkle it on the wound. • "I powdered it on until it was covered," Spievack recalled. • To his astonishment, every bit of his fingertip grew back. • "Your finger grew back?”, • "flesh, blood, vessels and nail?“ • "Four weeks," he answered.

21. Regenerating a finger and the extracellular matrix (4 of 4) • Alan worked with Steven Badylak of the University of Pittsburgh's McGowan Institute of Regenerative Medicine • The special powder was made up extracellular matrix: made from pig bladders. It is a mix of protein and connective tissue surgeons often use to repair tendons and it holds some of the secrets behind the emerging new science of regenerative medicine. • "It tells the body, start that process of tissue regrowth," said Badylak.

22. Regeneration and money (1 of 4) • In a [2008] clinical trial at Thomas Jefferson Hospital in Philadelphia, Dr. Patrick Shenotperformed a bladder transplant with an organ built with the patient's own cells. In a process developed by Dr. Atala[of Wake University], the patient's cells were grown in a lab, and then seeded on a biodegradable bladder-shaped scaffold. • Eight weeks later, with the scaffold infused with millions of regrown cells, it was transplanted into the patient. When the scaffold dissolves, Dr. Shenotsays, what's left will be a new, functioning organ. (CBS article by Wyatt Anderson)

23. Regeneration and money (2 of 4) • Corporate America, meanwhile, already believes regeneration will work. Investment capital has been pouring in to commercialize and mass produce custom-made body parts. • The Tengion Company has bought the license, built the factory, and is already making those bladders developed at Wake Forest that we told you about earlier.

24. Regeneration and money (3 of 4) • "We're actually building a very real business around a very real and compelling patient need," said Dr. Steven Nichtberger, Tengion's CEO. • Tengion believes regeneration will soon revolutionize transplant medicine. Transplant patients, instead of waiting years for a donated organ, will ship cells off to a lab and wait a few weeks to have their own re-grown.

25. Regeneration and money (4 of 4) • "I look at the patients who are on the waitlist for transplant," said Nichtberger. "I look at the opportunity we have to build bladders, to build vessels, to build kidneys. In regenerative medicine, I think it is similar to the semi-conductor industry of the 1980s, you don't know where it's going to go, but you know it's big.“(http://www.cbsnews.com/news/medicines-cutting-edge-re-growing-organs/)

26. The heart of the matter (1 of 6) • http://www.nature.com/news/tissue-engineering-how-to-build-a-heart-1.13327 • ( 5 min. video from 2013 article)

27. The heart of the matter (2 of 6) • Doris Taylor (director of regenerative medicine research at the Texas Heart Institute in Houston doesn't take it as an insult when people call her Dr Frankenstein. “It was actually one of the bigger compliments I've gotten,” she says — an affirmation that her research is pushing the boundaries of the possible.) She regularly harvests organs such as hearts and lungs from the newly dead, re-engineers them starting from the cells and attempts to bring them back to life in the hope that they might beat or breathe again in the living.

28. The heart of the matter (3 of 6) • The strategy is simple enough in principle. • First remove all the cells from a dead organ, it does not even have to be from a human [decellularize] • then take the protein scaffold left behind and repopulate it with stem cells immunologically matched to the patient in need [recellularize]

29. The heart of the matter (4 of 6) • Success with hollow organs (tracheas & bladders) • Problems with solid organs : • growing solid organs such as kidneys or lungs means getting dozens of cell types into exactly the right positions • simultaneously growing complete networks of blood vessels to keep them alive

30. The heart of the matter (5 of 6) • new organs must be sterile • able to grow if the patient is young, • and at least nominally able to repair themselves. • most importantly, they have to work — ideally, for a lifetime.

31. The heart of the matter (6 of 6) • The heart is the third most needed organ after the kidney and the liver, with a waiting list of about 3,500 in the United States alone, • but it poses extra challenges for transplantation and bioengineering. • The heart must beat constantly to pump some 7,000 litres of blood per day without a back-up. • http://www.nature.com/news/tissue-engineering-how-to-build-a-heart-1.13327

32. 2011 state-of-the-art heart rebuilding project • http://channel.nationalgeographic.com/channel/explorer/videos/the-re-animator/ (3 mins. 27 secs.)

33. 2013 heart research (1 of 3) • Decellularized mouse heart beats again after regenerating with human heart precursor cells (University of Pittsburgh researchers) • http://www.sciencedaily.com/releases/2013/08/130813112301.htm

34. 2013 heart research (2 of 3) • After 20 days of perfusion, the engineered heart tissues exhibit spontaneous contractions, generate mechanical force and are responsive to drugs. In addition, we observe that heart extracellular matrix promoted cardiomyocyte proliferation, differentiation and myofilament formation from the repopulated human multipotential cardiovascular progenitor cells.

35. 2013 heart research (3 of 3) • Repopulation of decellularized mouse heart with human induced pluripotent stem cell-derived cardiovascular progenitor cells by • Tung-Ying Lu • Bo Lin • Jong Kim • Mara Sullivan • KimimasaTobita • Guy Salams • & Lei Yang • http://www.nature.com/ncomms/2013/130813/ncomms3307/full/ncomms3307.html

36. All about skin • http://channel.nationalgeographic.com/channel/explorer/videos/the-skin-gun/ • JorgGerlach and his skin cell gun (3 mins. 30 secs.)

37. Feb. 2014 update on skin cell gun (1 of 4) • Question on Reddit (Feb. 2014) : What happened to the skin gun? • I actually work with in the Skin Gun Lab. It is true that not much press has come out since the National Geographic video. But we are still working with it and improving the process. We are still having occasional clinical trials and are working on getting FDA approval for the gun. I'm not sure what Kinds [sic] of specifics I can tell for legal reasons, but if you have specific questions I can try to answer them.

38. Feb. 2014 update on skin cell gun (2 of 4) • In addition, I'd like to say clarify something that bugs me about the national geographic video. • The police officer and narrator talk as if his wound was completely healed after 4 days. And this is not technically correct. The term that would be appropriate is that the wound was "re-epithelialized" which means that the wound has a thin layer of skin over it and is "dry." yet that layer of skin is extremely thin like paper and very fragile.

39. Feb. 2014 update on skin cell gun (3 of 4) • it takes weeks maybe months before the area is "completely" healed. • "re-epithelialization" in 4 days on a 2nd degree burn is quite good. those burns can take 2 weeks to heal on their own. depending on the depth of the burn. But to say the wound is completely healed in 4 days is not true.

40. Feb. 2014 update on skin cell gun (4 of 4) • I truly believe in the technology, but it should not be seen as a miracle, but rather an alternative to mesh grafting that leaves less scarring and can treat areas such as hands and face where mesh grafting is not possible. The analogy I love to use is that a mesh graft is similar to using sod and the skin gun is using grass seed. Hopefully as more research is done the process improves even on patients with less than optimal health. • http://www.reddit.com/r/askscience/comments/1y2e1r/what_happened_to_the_skin_gun/

41. How to build a beating heart • https://www.knowledge.ca/program/national-geographic-specials-how-build-beating-heart • Ear • Heart • Skin cell gun • And more

42. A return to the eye (1 of 6) • Hope for blind as scientists find stem cell reservoir in human eye • (Oct. 1, 2014 article for The Telegraph; http://www.telegraph.co.uk/science/science-news/11133622/Hope-for-blind-as-scientists-find-stem-cell-reservoir-in-human-eye.html)

43. Stem cells and the eye (2 of 6) • Researchers at the University of Southampton have discovered a reservoir of stem cells in an area of the eye called the corneal limbus. • And they have proven that, in the right environment, they can be transformed into photo-receptor cells which react to light.

44. Stem cells and the eye (3 of 6) • And researchers were amazed to find that the cells even existed in the eyes of a 97-year-old, opening up the possibility that the treatment could work for the elderly.

45. Stem cells and the eye (4 of 6) • University of Southampton: Centre for Human Development, Stem Cells and Regeneration • (http://www.southampton.ac.uk/chdscr/research/retinal_repair.page)

46. Stem cells and the eye (5 of 6) • Degenerative retinal diseases are the leading cause of untreatable blindness worldwide. There is currently no treatment for loss of light sensitive retinal cells (photoreceptors) or the cells which support their function known as retinal pigment epithelium (RPE) cells. Therefore cell based therapies are an attractive treatment option. We are interested in utilising stem cells, as potential cell sources for ocular transplantation.

47. Stem cells and the eye (6 of 6) • Our work includes isolating adult stem cells from the anterior eye, as well as other sources such as induced pluripotent stem cells. We then promote transdifferentiation towards retinal phenotypes including photoreceptor and RPE cells. • Paper: http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0108418 (Adult LimbalNeurosphere Cells: A Potential Autologous Cell Resource for Retinal Cell Generation)

48. Cells: immortal and stem (1 of 16) • An immortalized cell • proliferates indefinitely • can be naturally occurring or it could have been induced experimentally. • A cell becomes immortal due to a mutation

49. Cells: immortal and stem (2 of 16) • There are various immortal cell lines. • Some are normal cell lines - e.g. derived from stem cells • Other immortalised cell lines are the in vitro equivalent of cancerous cells

50. Cells: immortal and stem (3 of 16) • Cancer occurs when a somatic cell which normally cannot divide undergoes mutations that de-regulate the normal cell cycle controls and lead to uncontrolled proliferation • Immortalised cell lines have undergone similar mutations allowing a cell type which would normally not be able to divide to be proliferated in vitro