Stem cell & therapeutic cloning Lecture

1. Caption: This image depicts a colony of human embryonic stem cells grown over a period of 10 months in the absence of mouse feeder cells. The cell nuclei are stained green; the cell surface in red. Photo Credit: Courtesy Ren-He Xu/University of Wisconsin Caption: Cloned Mouse Description: A colony of embryonic stem cells. 10X. Image in the Public Domain. Description: (A) Human ESCs; (B) Neurons derived from Human ESCs. Images courtesy of Nissim Benvenisty.

3. Stem Cell History 1998 - Researchers first extract stem cells from human embryos 1999 - First Successful human transplant of insulin-making cells from cadavers 2001 - President Bush restricts federal funding for embryonic stem-cell research 2002 - Juvenile Diabetes Research Foundation International creates $20 million fund-raising effort to support stem-cell research 2003?? - California ok stem cell research–terminated! 2004 - Harvard researchers grow stem cells from embryos using private funding 2004 - Ballot measure for $3 Billion bond for stem cells

4. You may never read some stories. You may read some stories only once. You might read other stories over and over. All the cells in your body contain the same exact DNA. How your cells pick and choose the genes that will be expressed or “turned on” via transcription and translation is what makes cells different. These books represent different chromosomes in all your somatic cells. The stories within are the genes. The words are the genetic code. During development, most cells of the body “differentiate”.

5. The next few slides are pictures to remind you of the various stages of embryonic development=creation of life!

6. Embryonic development

8. A fertilized egg Notice the nuclei from the egg and sperm fusing

9. 4 day old embryos

10. 5 day old embryo at the blastocyst stage

11. Implantation

16. ESCs are pluripotent They retain the ability to develop into nearly any cell type

18. Generates every cell in the body including the placenta and extra-embryonic tissues Can form the entire human being Cannot form the entire human being Can generate every cell in the body except placenta and extra-embryonic tissues Become specific cell types; may or may not have plasticity Transdifferentiation?! WHAT?

19. STEM CELL (DEFINITION) A cell that has the ability to continuously divide and differentiate (develop) into various other kind(s) of cells/tissues Stem cell type Description Examples Totipotent Each cell can develop into a new individual Cells from early (1-3 days) embryos Pluripotent Cells can form any (over 200) cell types Some cells of blastocyst (5 to 14 days) Multipotent Cells differentiated, but can form a number of other tissues Fetal tissue, cord blood, and adult stem cells

24. How Big Is a Blastocyst?

28. In vitro fertilization Some procedures involved with IVF manually inject the sperm into the egg, and others simply allow fertilization to occur by adding the sperm to the egg in the lab setting.

32. Reprinted with permission of Do No Harm . Click on image for link to website.

33. Bone Marrow Stem Cells

35. Multipotent stem cell rich blood found in the umbilical cord has proven useful in treating the same types of health problems as those treated using bone marrow stem cells Umbilical Cord Blood Stem Cells In 2005, there were more than 1,400 cord blood transplantations in adults, according to NETCORD, an international network that coordinates umbilical cord blood banks Why would a parent consider this blood collection and why might it be considered to have advantages over bone marrow stem cells? The process to collect and store a child’s cord blood doesn’t come cheap. The company “Cord Blood Registry”, for example, charges and “initial fee” of $1975 and then it is $125 per year for storage.

36. Differences between embryonic stem cells and adult stem cells: -ESC can differentiate into any cell type (totipotent/pluripotent) while adult SC have already “committed” to a particular fate (multipotent). Some Challenges in Research: -Adult stem cells are often present in only minute quantities and can therefore be difficult to isolate and purify. -There is also evidence that they may not have the same capacity to multiply as embryonic stem cells do. -They do not have the development potential that a ESC -Finally, adult stem cells may contain more DNA abnormalities—caused by sunlight, toxins, and errors in making more DNA copies during the course of a lifetime. -These potential weaknesses might limit the usefulness of adult stem cells.

40. People with illnesses Parents of children with illnesses Physicians and scientists Research Institutes: NIH, Universities, Corporations and shareholders (those who might profit) Government (s) USA, California, UK, Australia, Czech Republic, South Korea Taxpayers Churches and clergy The cells themselves

41. Principle of Cell-based therapy Reprinted with permission from the Univ. of Kansas Medical Center. Click on image for link to originating website.

42. Cell-based therapy-Type 1 diabetes

43. Cell-based therapy- Spinal Cord Injury Differentiate (+ growth factors) START *Treatment may not work for the chronically paralyzed time Oligodendrocytes Clinical trials starting for treatment of spinal cord injury* in humans (after much data gathered using rats as an animal model) injured markedly recovered

44. Drug Development- Cancer Stem Cells Cell surface markers are a key difference. Reya, T., et al. Nature , 2001

47. Note myofibrils in finger–like projections attaching to microrod. Also note all myofibrils are highly oriented.

49. Cell-based therapy-Delivery chemotherapeutic agents

51. SCNT and Stem Cells Therapeutic Cloning or Nuclear Transplantation (SCNT = Somatic Cell Nuclear Transfer) Notice that UNLIKE reproductive cloning, all we are doing here is cloning embryonic cells and then coercing them to differentiate into specific cells the patient needs.

56. SCNT process (4) Ease the tip of the glass needle deep into the enucleated egg cell. Then, deposit the donor nucleus. Inject the nucleus (at arrow) from a donor cell into the enucleated egg cell. (Such a donor cell might be a skin cell from a disease sufferer whom doctors hope to treat using the patient’s own stem cells grown in culture)

59. (The cells may have their genes “corrected” before being transplanted to an individual.)

61. Asexual reproduction= Reproductive Cloning The Association of Reproductive Health Professionals

64. Cloning A Sheep (Roslin Institute http://www.roslin.ac.uk/library/)

66. Human Clones? Are there human clones? Yes – Identical Twins (Time 2.19.01) Learn from History- don’t repeat natures mistakes!

72. James Watson (L) and Francis Crick (R), and the model they built of the structure of DNA (based on the data garnered by Rosalind Franklin! ).

73. Hwang Woo-Suk was a professor and prominent researcher in the College of Veterinary Medicine at Seoul National University. In February 2004, Hwang and his team announced that they had successfully created embryonic stem cells with the somatic cell nuclear transfer method, and published their paper in Science (one of the most prestigious scientific journals there is) in March 2004. A second paper, published in May 2005, reported the creation of 11 stem cell lines that genetically matched nine patients with spinal cord injury, diabetes, and an immune system disorder.

78. That’s all folks!