© life_edu Lecture 25 Part Ib. Stem Cells - Therapy and Medical Research Issues in Biotechnology: The Way We Work With Life Dr. Albert P. Kausch life edu.us Medical Biotechnology

Issues in Biotechnology: Biotechnology, Our Society and Our Future OnCampus Live BCH 190, MIC 190, AFS 190, NRS 190, PLS 190 OnLine BCH 190 A Sweeping General Survey on Life and Biotechnology A Public Access College Course The University of Rhode Island Kimberly Nelson Issues in Biotechnology: The Way We Work With Life Dr. Albert P. Kausch life edu.us

© life_edu A Sweeping General Survey on Life and Biotechnology The University of Rhode Island Issues in Biotechnology: The Way We Work With Life Dr. Albert P. Kausch life edu.us BCH 190 Section II. The Applications of Biotechnology

Stem Cell Research What is involved? What is the potential? What are the downsides? Who decides?

What, for you, divides our thinking about stem cell research? (A) support for the research that may result in human cloning (B) uncertain research, not near clinical trials (C) lack of application, still unproven results (D) right to life issues (E) lack of information on the topic

Do you support federal funding for embryonic stem cell research? (A) yes (B) no (C) yes, but with restrictions

Therapeutic vs Reproductive Cloning Embryonic Stem Cells vs Adult Stem Cells vs iPS Cells

Stem Cell Research Obama lifts the ban of Federal funding for Stem Cell Research New research will bring innovation

Stem Cell Basics

Embryonic, Adult and Induced Pluripotent Stem Cells

Stem Cell Basics What is a Stem Cell? What Are the Different Types of Stem Cells? Stem Cell Therapy: How will this be used? What is the progress? What are the controversies?

Characteristics of Stem Cells: A capacity for extensive self-renewal Developmental potential (The ability to differentiate) The type of stem cell is defined both by its origin (site, species, age) and by its differentiation abilities What is a stem cell?

Examples of types of stem cells based on age of organism at time of derivation Embryonic Fetal Newborn Adult iPSc

Stem cells give rise to specialized cells residing in organs Stem Cells

Stem Cell Self-renewal Symmetric Cell Division (Proliferation) 1 stem cell 2 stem cells

Stem Cell Differentiation Asymmetric Cell Division (Proliferation) 1 stem cell 1 stem cell + 1 differentiated cell

Stem Cells Totipotency

Types of Stem Cells Embryonic Stem Cells (ES Cells)--Derived from early embryos (blastocysts) ES cells are totipotent--they can form all the cell-types in an adult ES cells are embryonic stem cells and immortal (unlimited proliferation in dishes) hES cells are Human embryonic stem cells Adult Stem Cells - Derived from pluripotent adult tissues (i.e. bone marrow transplants) IPS Cells - Induced pluripotent cells are derived from adult tissues and genetically modified with genes that override the cell division pathways to become stem cells capable of then differentiating

Embryonic Stem Cells (ES cells)

Preimplantation Development Morula Blastocyst Zygote ICM Extraembryonic progenitors

Preimplantation Development Morula Blastocyst Zygote ICM Extraembryonic progenitors

Derivation of Embryonic Stem Cells Morula Zygote Blastocyst ES cells

Totipotency of ES cells Zygote Morula Blastocyst ES cells

How embryonic stem cell lines are produced Dissociate blastocyst into single cells Grow in the presence of growth factors Only rarely does a single cell continue to grow indefinitely

How embryonic stem cell lines are validated Ectoderm Skin Neurons Mesoderm Muscle Blood Bone Endoderm Pancreas Lung Liver Grow cells under different conditions to stimulate differentiation into multiple different cell types

Immuno-rejection

Solutions to immuno-rejection Personalized Stem Cells Somatic cell nuclear transfer Unfertilized egg cell Remove nucleus from the egg cell and replace it with that of the patient Use newly formed stem cells for therapeutic purposes Specific to the patient In addition, ES cells can be genetically engineered Cell from patient These cells can also be genetically engineered

Promoter Coding Sequence Terminator Stem Cells can be Genetically Modified Genetic constructs could be used in stem cell therapy Controlled expression Cell and developmentally specific Your favorite gene Insulin Anti-cancer Tay-sachs many others Stop transcription Message stability

Nuclear Transfer Stem Cell Therapy Advantages: No Rejection (Complete tissue- type match) Complete Cures Versatile-Genetic Modification Disadvantages: Controversial Expensive Requires supply of oocytes

ES Cell Technology is Well Established in Mice Morula Blastocyst Zygote ICM Extraembryonic progenitors Use of ES technology is well established in the mouse model ES cells from mice can be made genetically engineered and established in recipients

The same Technology Could be Applied to Humans Morula Blastocyst Zygote ICM Extraembryonic progenitors The same technology could be used to clone a human Therapeutic vs Reproductive Cloning Embryonic Stem Cells vs Adult Stem Cells vs iPS Cells

Adult Stem Cells

Types of Stem Cells: Adult Stem Cells--A small subpopulation of specialized stem cells present within some adult organs capable of self renewal Adult stem cells are pleuripotent--They have more limited potential Adult stem cells cannot divide indefinitely

Adult Stem Cells pluripotency

Hematopoietic stem cells are capable of differentiation into all mature peripheral blood cells Liver stem cells are capable of differentiation into hepatocytes and cholangiocytes Neural stem cells can differentiate into neurons, astroglia and oligodendrocytes Some examples of types of stem cells based on their differentiation abilities

Adult Stem Cell Therapy Steps: 1. Biopsy 2. In vitro culture of Adult Stem Cells 3. In vitro expansion or differentiation (If needed) 4. Genetic modification (as needed) 5. Engraftment

Bone Marrow Transplantation Plasticity: The ability of cells to change developmental fate GFP Donor Bone Marrow (Contains Hematopoetic Stem Cells) Engraftment Repopulation of “Lethally” Irradiated Host (Lymphoid + Myeloid) Genetic Modification Adult stem cell differentiation has been demonstrated

Adult Stem Cell Therapy Advantages: No Immuno-Rejection (if from self) Not Controversial Disadvantages: Many organs do not contain adult stem cells Stem cells may be absent or defective in the affected organs Use of Plasticity and transdifferentiation is unproven

Adult Stem Cells Self Renewal (symmetric cell division)

Adult Stem Cells Self Renewal Activation, Proliferation (asymmetric cell division)

Adult Organs Bone Marrow/ Blood Skin Liver Pancreas Intestine Testes Brain ( but limited) Organs With Adult Stem Cells More to come? Probably! And these can be genetically modified to suit a specific disease or disorder

Question about Plasticity Plasticity: The ability of cells to change developmental fate GFP Donor Bone Marrow (Contains Hematopoetic Stem Cells) Engraftment Repopulation of “Lethally” Irradiated Host (Lymphoid + Myeloid) + Neurons (a few!)

True Plasticity GFP + Donor Bone Marrow (Contains Hematopoetic Stem Cells) Engrafted Cells (Lymphoid + Myeloid) Neurons (a few!)

Induced Pluripotent Stem Cells (iPS)

Induced pluripotent stem cells, commonly abbreviated as iPS cells or iPSc, are a type of pluripotent stem cell artificially derived from a non-pluripotent cell, typically an adult somatic cell, by inducing a “forced” expression of certain genes that regulate the cell division cycle thus making them immortal. Induced Pluripotent Stem Cells are believed to be identical to natural pluripotent stem cells, such as embryonic stem cells in many respects, such as the expression of certain stem cell genes and proteins, chromatin methylation patterns, doubling time, embryo formation, teratoma formation, viable chimera formation, and potency and differentiability, but the full extent of their relation to natural pluripotent stem cells is still being assessed. IPScs were first produced in 2006 from mouse cells and in 2007 from human cells. This has been cited as an important advancement in stem cell research, as it may allow researchers to obtain pluripotent stem cells, which are important in research and potentially have therapeutic uses, without the controversial use of embryos.

Induced Pluripotent Stem Cell Basics A scheme of the generation of induced pluripotent stem (iPS) cells. (1) Isolate and culture donor cells. (2) Transfect stem cell-associated genes into the cells by viral vectors. Red cells indicate the cells expressing the exogenous genes. (3) Harvest and culture the cells according to ES cell culture, using mitotically inactivated feeder cells (4) A small subset of the transfected cells become iPS cells and generate ES-like colonies.feeder cells INDUCED PLURIPOTENT Stem Cell Lines Derived from Human Somatic Cells Junying Yu 1*, Maxim A. Vodyanik 2, Kim Smuga-Otto 1, Jessica Antosiewicz-Bourget 1, Jennifer L Frane 3, Shulan Tian 4, Jeff Nie 4, Gudrun A. Jonsdottir 4, Victor Ruotti 4, Ron Stewart 4, Igor I. Slukvin 5, James A. Thomson 6 Science DOI: /science

Stem Cell Therapy: Recent Progress

Stem Cell Therapy

Human Degenerative Disorders: Many diseases are characterized by organ failure or cell loss: Degenerative disease states (unknown etiology or autoimmune disease) Age-related degenerative disease Cell or organ destruction due to genetic or infectious disease

Stem Cell Therapy: Stem Cell Therapy is the replenishment of cells or organs lost to degenerative disease or loss of function with new cells derived from stem cells Stem Cell Therapy is Experimental (Except Bone Marrow Transplantation) Stem Cell therapy should be free of pharmaceutical toxicity Cellular Pharmaceutical

Stem Cell Therapy Stem Cell Therapy offers a method to treat degenerative disorders A Cellular Pharmaceutical Diseases amenable to Stem Cell Therapy  Diabetes  Spinal chord injury  Alzheimer’s Disease  Parkinson’s Disease  Heart Disease  Many more…..but not overnight

Stem Cell Therapy Established Stem Cell Therapies: Bone Marrow Transplantation--Restores blood system with hematopoetic stem cells (1950s) (many variations) Simplest Diseases to cure next involve secretory cells:  Parkinson’s Disease--Dopamine Secretion  Type I Diabetes--Insulin Secretion (Type II is difficult due to insulin insensitivity)

ES cell Therapy (Parkinson’s Disease) ES cells Dopaminergic Neurons Engraftment Dopamine Production Restored in vitro: in vivo:

Parkinson’s Disease-Experimental Therapy From Lindvall et al., Nat. Med. 2004, S42-S50.

Stem Cell Therapy Criticisms and Controversies

Embryogenic Stem Cell Criticisms I. Destruction of human embryos II. A slippery slope to: Human Genetic Engineering Human cloning

Somatic cell nuclear transfer techniques are the same as those that could be applied to cloning Unfertilized egg cell Remove nucleus from the egg cell and replace it with that of the patient Cell from patient These cells can also be genetically engineered Use newly formed stem cells for therapeutic purposes Specific to the patient In addition, ES cells can be genetically engineered

Where is DNA in a cell? All of the information for each protein in a cell is in it’s DNA All of the information that codes for a complete organism is in DNA

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Credits Credits Lectures by: Edited by: Video Produced by: Thank You to The University of Rhode Island Thank You to The University of Rhode Island and all of the students of Issues in and all of the students of Issues in Biotechnology over the years Biotechnology over the years Dr. Albert Kausch and Kimberly Nelson Thaddeus Weaver