ALL ABOUT STEM CELLS

Compiled by Dr. Taruna Anand, Scientist

VTC, NRC Equine, Hisar (Haryana)-India

Dr. Dharmendra Kumar, Scientist CIRB, Hisar (Haryana)- India

What are Stem Cells?

The raw material from which all of the bodys mature, differentiated cells are made.
Unspecialized cells Give rise to more than 250 specialized cells in the body-brain cells, nerve cells, heart cells, pancreatic cells, etc. Serve as the bodys repair system Renew itself Replenish other cells

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

Whats So Special About Stem Cells?

They have the potential to replace cells/tissue that has been damaged or destroyed by severe illnesses.

They can replicate themselves over and over for a very long time.
Understanding how stem cells develop into healthy and diseased cells will assist the search for cures.

Two Kinds of Stem Cells

Embryonic stem cells are capable of developing into all the cell types of the body Adult stem cells are less versatile and more difficult to identify, isolate, and purify

Stem Cell Characteristics

Blank cells (unspecialized) Capable of dividing and renewing themselves for long periods of time (proliferation and renewal) Have the potential to give rise to specialized cell types (differentiation)

Stem Cell Differentiation

Where Are Early Stem Cells Found?

Who did it first?

In 1998, U. Wisconsin research team isolated stem cells from IVF-blastocysts

Stem Cell Cultivation

What are the sources of Mature Stem Cells?

Mature Body Tissues

Umbilical Cord & Placenta SC Research

Isolated immediately following birth Whartons Jelly showing promise as a source More flexibility = some pluripotent characteristics Research is limited but growing

Human Stem Cell Production

Stem cells can be manipulated outside the body for therapies

assist wound healing grow new cells correct defective gene(s) gene therapy off-the-shelf, pre-fabricated body parts?

Stem cells
renewable source of replacement cells and tissues to treat diseases

Stem cell therapy to treat Parkinson's disease in human

Step 1 : Define the problem

Step 2 : Finding the right type of stem cell

Stem cells treat cardio vascular disease

Adult stem cells - administered intravenously - can regenerate damaged heart muscles

Bone marrow stem cells to treat leukemia

- Bone marrow stem cells introduced into patients blood stream - Produce new healthy leukocytes to replace abnormal cells

Stem cells made to produce insulin cure Diabetes

Stem cells from hair can grow into skin Skin replacement

Spinal cord injuries Multiple sclerosis Alzheimers disease

Stem cell therapy in animals

 Stem cells treat heart attack in animals

In pigs, bone marrow stem cells repaired damaged heart muscle

Treatment of arthritis, tendon and ligament injuries

Fat derived stem cells heal injuries in dogs and horses

Canine and equine applications

200 dogs and 3000 horses successfully treated for Tendon injury: acute and chronic Ligament injury: acute and chronic Joints: osteoarthritis

Applications of stem cells in farm animals

For cloning For the generation of transgenic animals For rapid screening of chemicals and drug discovery For obtaining germ cells to treat infertility For examining complex processes such as genomic imprinting For studies on early embryonic development For gene targeting

P R O D U C T I O N o F ES C E L L S

Blastocysts

Development of feeder layers Isolation of ICM cells

Culture of ICM cells on feeder layers for production of ES cell lines

Characterization of putative ES cells for expression of different markers such as:

Immunohistochemical Surface Markers Transcription based Markers

Immunosurgery

Intact blastocyst Culture

Methods for isolation of ES cells from embryos

Mechanical Isolation

Enzymatic Digestion

Isolation of ES cells

Immunosurgery

Mechanical isolation

Feeder layer
Layer of non-dividing cells which provides nutrients and necessary growth factors Prevents differentiation of ES cell as it secretes LIF Cells are inactivated by:

- irradiation
mitomycin-C

Development of feeder layer

Primary culture Fetus

Sub-dermal tissue is minced into small pieces in DPBS

Treatment with trypsin-EDTA and neutralization with DMEM+FBS Centrifugation

Cells cultured in flasks at 38.5C in CO2 incubator to confluency Subcultured (P5) Harvested cells were cryopreserved Confluent monolayer at passage-5 Frozen aliquots thawed and grown to confluency

Treatment with mitomycin C (10g/ml-2hr) prior to usage

Embryonic stem cells grow in tightly bound colonies

Adult stem cells

The use of embryonic and foetal stem cells is controversial - many people believe that the early embryo is sacrosanct. An alternative might be to find a source of stem cells in adults. Scientists have found that certain bone marrow cells behave very like embryonic cells and can be transformed into different types of cell.

Culture of Umblical Cord Blood Cells

Ways to identify stem cells

Pluripotent nature Morphology Longevity Genetic stability Epigenetic status

Expression of markers

Differentiation to embryoid bodies

Directed differentiation

Ethics in stem cell research

What is ethics ?
Ethics refers to a set of moral principles that govern the appropriate conduct of an individual or group. Ethics speaks to how we ought to live, that is, how we ought to treat others and how we ought to run or manage our own lives.

Basic Ethical Problem

Enjoins the prevention or alleviation of suffering. Enjoins us to respect the value of human life.

Issues regarding stem cell research

There are several types of issues to consider as we think about stem cell research. Ethical issues are those that ask us to consider the potential moral outcomes of stem cell technologies. Legal issues require researchers and the public to help policymakers decide whether and how stem cell technologies should be regulated by the government. Social issues involve the impact technologies on society as a whole. of stem cell

The Value of the Embryo

There are three broad schools of thought about the moral status of the early embryo.

1)

2)

3)

Moral status of embryo is equal to that of a human adult. Embryos potential for personhood develops as it grows. Personhood is required for moral status.

Conti.
The use of stem cells in regenerative medicine potentially could cure some currently untreatable forms of disability. Why then, would some people with disabilities be opposed to ESC research?
Which view do you accept of when we should accord a human Embryo or foetus special moral status?

There is general agreement in the community that brain death in a human being defines death What features of an organism at the beginning of its existence might be relevant in deciding when human life begins?

Are pro-life leaders ignoring the real problems?

World stem cell map

Permissive
Australia, Belgium, China, India, Israel, Japan, Singapore, South Korea, Sweden, the United Kingdom

Flexible
Brazil, Canada, France, Iran, South Africa, Spain, The Netherlands, Taiwan

Restricted policy
Austria, Ireland, Norway, Poland, Germany, Italy, and the United States

Draft guidelines for stem cell research in India

DBT & ICMR

Permissible Adult Umbilical cord blood

Restricted

Prohibited Human cloning

ESCs

Regulations
ESC Research Donors consent
Cord blood banks Drug Control General of India Penalty for misconduct - Monetary fine, Jail

Pregnancy cant be attempted for donating foetal tissues

National Apex Committee for Stem cell Research and Therapy

Institutional Committee for Stem cell Research and Therapy

For Further Investigation

Form an Ethics Committee

Lawyer Various religious representatives

Lay community representatives

Disability representatives Scientists

Stem cells
Are the magic cells capable of regeneration

Have the potential cells/tissue

to

replace

damaged

Can be used for the animal welfare too Can be cultured in the laboratory preserved as stem cell lines Hold great promises for disease treatment and