STEM CELL CULTURE TECHNOLOGY Importance of Stem Cell

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STEM CELL CULTURE TECHNOLOGY

STEM CELL CULTURE TECHNOLOGY

Importance of Stem Cell Research 12/3/2020 Dr. Hariom Yadav

Importance of Stem Cell Research 12/3/2020 Dr. Hariom Yadav

Stem Cell History 1998 - Researchers first extract stem cells from human embryos 1999

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 2002 - California ok stem cell research 2004 - Harvard researchers grow stem cells from embryos using private funding 2004 - Ballot measure for $3 Billion bond for stem cells 12/3/2020 Dr. Hariom Yadav

Stem Cell – Definition • A cell that has the ability to continuously divide

Stem Cell – Definition • A cell that has the ability to continuously divide and differentiate (develop) into various other kind(s) of cells/tissues 12/3/2020 Dr. Hariom Yadav

Stem Cell Characteristics Ø ‘Blank cells’ (unspecialized) Ø Capable of dividing and renewing themselves

Stem Cell Characteristics Ø ‘Blank cells’ (unspecialized) Ø Capable of dividing and renewing themselves for long periods of time (proliferation and renewal) Ø Have the potential to give rise to specialized cell types (differentiation) 12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

Kinds of Stem Cells Stem cell type Totipotent Pluripotent Multipotent 12/3/2020 Description Each cell

Kinds of Stem Cells Stem cell type Totipotent Pluripotent Multipotent 12/3/2020 Description Each cell can develop into a new individual Examples Cells from early (13 days) embryos Some cells of Cells can form any (over blastocyst (5 to 14 200) cell types days) Cells differentiated, but Fetal tissue, cord can form a number of blood, and adult other tissues stem cells Dr. Hariom Yadav

Stem Cell Differentiation 12/3/2020 Princeton University Dr. Hariom Yadav

Stem Cell Differentiation 12/3/2020 Princeton University Dr. Hariom Yadav

Kinds of Stem Cells Embryonic stem cells come from a five to six-day-old embryo.

Kinds of Stem Cells Embryonic stem cells come from a five to six-day-old embryo. They have the ability to form virtually any type of cell found in the human body. Embryonic germ cells are derived from the part of a human embryo or foetus that will ultimately produce eggs or sperm (gametes). Adult stem cells are undifferentiated cells found among specialised or differentiated cells in a tissue or organ after birth. Based on current research they appear to have a more restricted ability to produce different cell types and to self-renew. 12/3/2020 Dr. Hariom Yadav

Blastocyst Diagram 12/3/2020 Princeton University Dr. Hariom Yadav

Blastocyst Diagram 12/3/2020 Princeton University Dr. Hariom Yadav

Sexual Reproduction 12/3/2020 The Association of Reproductive Health Professionals Dr. Hariom Yadav

Sexual Reproduction 12/3/2020 The Association of Reproductive Health Professionals Dr. Hariom Yadav

Stages of Embryogenesis Day 1 Fertilized egg 12/3/2020 Day 2 2 -cell embryo Day

Stages of Embryogenesis Day 1 Fertilized egg 12/3/2020 Day 2 2 -cell embryo Day 11 -14 Dr. Hariom Yadav Tissue Differentiation Day 3 -4 Multi-cell embryo Day 5 -6 Blastocyst

Derivation and Use of Embryonic Stem Cell Lines Isolate inner cell mass (destroys embryo)

Derivation and Use of Embryonic Stem Cell Lines Isolate inner cell mass (destroys embryo) Outer cells (forms placenta) Inner cells (forms fetus) Day 5 -6 Blastocyst 12/3/2020 “Special sauce” (largely unknown) Dr. Hariom Yadav Kidney Culture cells Heart muscle Heart repaired

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

Embryonic stem cells 12/3/2020 Dr. Hariom Yadav

Embryonic stem cells 12/3/2020 Dr. Hariom Yadav

Adult stem cells 12/3/2020 Dr. Hariom Yadav

Adult stem cells 12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

Applications Ø Disease • Diabetes, Spinal cord injury, Parkinson’s disease, heart disease Ø Genetic

Applications Ø Disease • Diabetes, Spinal cord injury, Parkinson’s disease, heart disease Ø Genetic based Disease • Cystic fibrosis, Huntington’s 12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

Unknowns in Stem Cell/Cloning Research It is uncertain that human embryonic stem cells in

Unknowns in Stem Cell/Cloning Research It is uncertain that human embryonic stem cells in vitro can give rise to all the different cell types of the adult body. • It is unknown if stem cells cultured in vitro (apart from the embryo) will function as the cells do when they are part of the developing embryo • 12/3/2020 Dr. Hariom Yadav

Challenges to Stem Cell/Cloning Research Stem cells need to be differentiated to the appropriate

Challenges to Stem Cell/Cloning Research Stem cells need to be differentiated to the appropriate cell type(s) before they can be used clinically. • Recently, abnormalities in chromosome number and structure were found in three human ESC lines. • 12/3/2020 Dr. Hariom Yadav

Challenges to Stem Cell/Cloning Research Stem cell development or proliferation must be controlled once

Challenges to Stem Cell/Cloning Research Stem cell development or proliferation must be controlled once placed into patients. • Possibility of rejection of stem cell transplants as foreign tissues is very high. • 12/3/2020 Dr. Hariom Yadav

Challenges to Stem Cell/Cloning Research Contamination by viruses, bacteria, fungi, and Mycoplasma possible. •

Challenges to Stem Cell/Cloning Research Contamination by viruses, bacteria, fungi, and Mycoplasma possible. • The use of mouse “feeder” cells to grow ESC could result in problems due to xenotransplantation (complicating FDA requirements for clinical use). • 12/3/2020 Dr. Hariom Yadav

Stem cell and cloning Immune rejection Somatic cell nuclear transfer 12/3/2020 Dr. Hariom Yadav

Stem cell and cloning Immune rejection Somatic cell nuclear transfer 12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

12/3/2020 Dr. Hariom Yadav

Thanks 12/3/2020 (Time 2. 19. 01) Dr. Hariom Yadav

Thanks 12/3/2020 (Time 2. 19. 01) Dr. Hariom Yadav

References: 1. Dev. Biol- Verma and Agarwal 2. Dev. Biol- Wolpert 3. Dev. Biol.

References: 1. Dev. Biol- Verma and Agarwal 2. Dev. Biol- Wolpert 3. Dev. Biol. - Gilbert 4. Embryology- Ballinsky 5. Wikipedia 6. Freely available resources on internet 12/3/2020 Dr. Hariom Yadav