Stem cell

Therapy, introduction, applications

1. ADVANCES IN MICROBIOLOGY STEM CELL THERAPY

2. OVERVIEW OF STEM CELLS Stem cell biology Requirements & Potency definition Types of stem cell Stem cell collection Cryopreservation of stem cell Stem cell banking Stem cell therapy-disease treatment Benefits& limitation of stem cell Application of stem cell

3. STEM CELL BIOLOGY The science of stem cells is known as stem cell biology. It has been one of the fascinating areas of biology today provides cell-based therapies to treat reparative diseases. STEM CELLS The undifferentiated animal cells that are capable of proliferation and differentiation to do specialized functions are called stem cells. 1. Actual stem cells 2. Potential stem cells The body is made up of about 200 different kinds of specialised cells such as muscle cells, nerve cells, fat cells and skin cells. All cells in the body come from stem cells ( Stem cells are the building blocks of the blood and immune systems. Stem cells can self-renew to make more stem cells differentiate into a specialized cell type WHY ARE STEM CELLS SPECIAL?

4. CHARACTERISTICS OF STEM CELLS Stem cells are cells are capable of continuous proliferation for several months. Sub-culturing the stem cells for more than six months and observing the culture under a microscope for a clump formation. The stem cells do not form clumps. They have identical chromosomes as in diploid cells of the adult organisms. They can be sub-cultured, stored in liquid nitrogen . They are pluripotent . This is confirmed by allowing the ESCs. They have no tumour inducing property . When the ESCs are injected into immunosuppressed mouse, they do not induce tumours in that mouse. POTENCY DEFINITIONS Pluripotent: Stem cells that can differentiate into any tissue type except for placenta tissue. Multipotent: Stem cells that can differentiate into multiple cells in a closely related family of cells.

5. MULTPOTENT PLURIPOTENT blood stem cell found in bone marrow MULTIPOTENT only specialized types of blood cell: red blood cells, white blood cells, platelets differentiation Stem cells that can become many types of cells in the body are called pluripotent Stem cells that can become only a few types of cells are called Multipotent Tissue stem cells (Multipotent) Embryonic stem cells (pluripotent)

6. TYPES OF STEM CELLS 1. EMBRYONIC STEM CELLS 2. ADULT STEM CELLS EMBRYONIC STEM CELLS The undifferentiated cells isolated from embryos at blastocyst stage are called (ESCs). These cells are capable of continuous proliferation and differentiation, leading to the development of many different cell types in the adult. Hence, they are said to be Pluripotent. ESCs are found as the inner cell mass in the blastocyst. ESCs (Germ layer) Endoderm Ectoderm Mesoderm (Thymus,thyroid,lung, (Skin,nerves,head, (Bonemarrow,blood liver,pancreas, gastro eyes, ears) cells,muscles, heart, -intestinal tract) blood vessels)

7. EMBRYONIC STEM (ES) CELL CULTURE feeder layer Blastocyst embryonic stem cells taken from the inner cell mass outer layer of cells = ‘trophectoderm ’ cells inside = ‘inner cell mass ’ culture in the lab to grow more cells fluid with nutrients embryonic stem cell PLURIPOTENT differentiation A ll possible types of specialized cells

8. ADULT STEM CELLS The stem cells isolated from fully developed organs are called adult stem cells (ASCs). They can be isolated from liver,skin,heart, bonemarrow,blood stream,muscles, pancreas. In these organs, adult stem cells are found in discrete populations. If a part of these organs are injured, the adult stem cells regenerate to heal up the injury. Thus they regenerate tissues lost due to injury, tear or disease. FEATURES OF ASCs: They are tissue specific stem cells. Each ASC type gives only a particular cell type present in the organ from which it was isolated. Stem cell obtained from one organ may give rise to a cell type of yet other organ. This is called plasticity.

9. STEM CELL COLLECTION Peripheral Blood Stem Cell Collection Bone Marrow Harvest: Stem cells can be collected from the blood. This technique does not require surgery. It conventional bone marrow harvest. Prior to the collection, the donor is given a medication to promote the growth and release of stem cells from the bone into the blood. The stem cells are then collected using a special machine called a Cell Separator.. Stem cells are generally collected using this method here at the Leukemia. Stem cells can be collected directly from the bone marrow spaces, most often from the pelvic bones. Several puncture sites are made along the bone and the cells are removed using a needle. This procedure is known as a  bone marrow harvest . Note that this technique is used less often here at the Leukemia.

10. CRYOPRESERVATION Cell lines (SC) have been selected for long time storage at -196ºC. The selected cell lines are subjected to various test such as Cytological, biochemical, physiological & immunological tests. COS media 3 contains 100µg/ml Streptomycin sulfate, 20% Calf serum, 100 units/ml Penicillin-G, 1% L-Glutamine& Dulbecco’s Modified Eagle Medium per litre. During preservation, cells are kept on Solid carbon dioxide (-79ºC) or Liquid Nitrogen (-196ºC).

11. STEM CELL (CORD BLOOD)BANKING An   amniotic stem cell bank  is a facility that stores stem cells derived from amniotic fluid  for future use. The first private amniotic stem cell bank ( life cell ) in the US was opened by  Biocell Center in October 2009 in Medford. Cord blood banking newborn's umbilical cord and placenta and storing it for future medical use. Cord blood contains potentially lifesaving cells called stem cells. Life cell-largest private bank 2004 in Chennai. Cord blood( bio-insurance ) is a rich source of blood stem cells. Stem cells are also found in bone marrow, human embryos, fetal tissue, hair follicles, baby teeth, fat, circulating blood, and muscle. Every part of the human body contains some stem cells, but most are not a rich enough source to be harvested for therapeutic applications.

12. HUMAN EMBRYONIC STEM CELLS The undifferentiated cells present in the inner cell mass of human blastocyst embryo are called human embryonic stem cells (HESCs). (HESCs) have been used in cell-based therapies to cure several diseases in man. Stem cell research since 1998. The HESCs can be used to prepare translating cells to cure Parkinson’s disease, diabetes, heart disease, vision loss and hearing loss, autism, Alzheimer's diseases,etc. Cord blood stem cells have been used successfully to treat more than 70 different diseases. Researches on stem cells to treat these diseases are progressing every year towards success.

13. APPLICATIONS IN DISEASE TREATMENT LEUKEMIA TYPE 1 DIABETES HEART DISEASE(MUSCLE REPAIR) PARKINSON’S DISEASE(PD)

14. LEUKEMIA Leukemia is cancer of the blood or bone marrow(which produce blood cells), person suffer abnormal production of blood cells [Leukocytes(WBC)]. FUNCTION OF THE BONEMARROW: Marrow-large bones of adults produces blood cells,4% our total body weight consists of bone marrow. TWO TYPES OF BONE MARROW: Red marrow: WBC - Fight disease RBC - Carry oxygen PLATLETS - Essential for blood clotting Yellow marrow : Inside middle section of the long bones.

15. TYPES OF LEUKEMIA : Acute Lymphocytic Leukemia Chronic Lymphocytic Leukemia Acute Myelogenous Leukemia Chronic Myelogenous Leukemia SYMPTOMS OF LEUKEMIA: Blood clotting is poor, affected immune system, anemia, fever, nausea, night sweats, tiredness ,headache & weight loss. CAUSES OF LEUKEMIA: Artificial ionizing radiation,HIV, hair dyes, genetic predisposition(family history). TREATMENTS: Chemotherapy , radiation therapy, stem cell transplant .

16. STEM CELL TRANSPLANTATION FOR LEUKEMIA: It is also called Hematopoietic progenitor cell transplantation. S.C collect from the peripheral blood& umbilical cord blood. STEM CELL TRANSPLANT:  ( BONE MARROW) A stem cell transplant is a procedure to replace diseased bone marrow with healthy bone marrow. Before a stem cell transplant, receive high doses of chemotherapy or radiation therapy to destroy diseased bone marrow. Then receive an infusion of blood-forming stem cells that help to rebuild bone marrow. May receive stem cells from a donor, or in some cases may be able to use your own stem cells. A stem cell transplant is very similar to a bone marrow transplant. TYPES: Autologous stem cell transplant Allogeneic stem cell transplant

17. AUTOLOGOUS STEM CELL TRANSPLANT ALLOGENIC STEM CELL TRANSPLANT Stem cell collected from the patients blood Harvested Frozen Stored until needed (Radiation & Chemotherapy) Destory the cancer cells Stem cell taken from matching donor Relative& family Umbilical cord blood (Sibling) Donor’s stem cells are the right match ( HLA test), compare the patient’s blood & tissue type with blood samples from the donor.

18. STEM CELL TRANSPLANTATION FOR LEUKEMIA

19. DIABETES Diabetes mellitus  ( DM ): It is simply known as   diabetes , is a group of metabolic diseases in which there are high blood sugar levels over a prolonged period.  The normal cells of pancreas that produce insulin are destroyed by the patients own immune system, leading to diabetes in humans. TYPES OF DIABETES There are three main types of diabetes mellitus: Type 1 DM: Results from the body's failure to produce enough insulin. This form was previously referred to as "insulin-dependent diabetes mellitus" (IDDM). Is an autoimmune disease in which the body attacks beta cells in the pancreas that secrete the important hormone insulin.

20. Type 2 DM :  Insulin resistance, cells fail to respond to insulin properly. As the disease progresses a lack of insulin may also develop. This form was previously referred to as "non insulin-dependent diabetes mellitus" (NIDDM) or "adult-onset diabetes". The primary cause is excessive body weight and not enough exercise.(family history). Gestational diabetes : Is the third main form and occurs when pregnant women without a previous history of diabetes develop a high blood glucose level. SYMPTOMS: This high blood sugar produces the symptoms of frequent urination, increased thirst, and increased hunger. Untreated, diabetes can cause many complications. Acute complications include coma.  heart disease, stroke, kidney failure, foot ulcers and damage to the eyes.

21. PREVENTION AND TREATMENT : Involves a healthy diet, physical exercise, not using tobacco and being a normal body weight. Blood pressure control and proper foot care are also important for people with the disease. Type 1 -diabetes must be managed with insulin injections. Type 2- diabetes may be treated with medications with or without insulin. Insulin and some oral medications can cause low blood sugar.  Gestational diabetes-  usually resolves after the birth of the baby.

22. STEM CELL BASED THERAPY Isolated islet cells from pancreas of foetus, purified and introduced them into pancreas of patients suffering Type-I diabetes. It increased the insulin concentration for a few days and thereafter the insulin concentration decreased slowly.(transplanted cells remained in the pancreas). The gene PDX1- insulin producing gene in cells . Isolated PDX1 gene& introduced into human embryonic stem cells to differentiate them into insulin secreting cells ( β -cells of pancreas). Cells were introduced into pancreas using a laparoscopic device. The introduced cells formed of islets like groups of cells and produced insulin continuously in the patients.

23. This technique has been tried to cure diabetes in clinical laboratories

24. PARKINSON’S DISEASE (PD) Parkinson's disease is a progressive nervous system disorder that affects the person moves, including they speak and write. The PD is a neurodegenerative disorder seen in 2% of human population over 65 years of age. It is due to progressive degeneration and loss of dopamine -producing neurons. TREATMENT: Embryonic stem cell transplantation was used. Lab-differentiated ESCs into dopamine, producing neurons for transplantation in patient suffering from PD. The PD cannot be treated with known drugs. New Technique Could Treat Parkinson’s Using Patient’s Own Brain Cells Scientists have successfully implanted nerve stem cells into the brain of a monkey, targeting the area destroyed by Parkinson’s disease .

25. EMBRYONIC STEM CELL TRANSPLANTATION Nurr 1 gene was introduced into mouse ESCs to differentiate them into dopamine producing neurons. Then the differentiated neurons are transplanted into the brain of the Parkinson's model mouse. The differentiated neurons reinnervated the brain, released dopamine and improved the motor function. Attempts to differentiate dopamine producing neurons from HESCs for transplantation in Parkinson's patients. Neurotransplantation for Parkinson's disease available in the near future.

26. STEM CELL TREATMENT Abstract Parkinson’s disease (PD) is a neurodegenerative disorder caused by a progressive degeneration of the midbrain dopamine (DA) neurons in the substantia nigra pars compacta (SNc) that predominantly affects the ventral population projecting to the dorsal striatum and leads to a gradual dysfunction of the motor system. There is currently no cure for PD. Pharmacological and surgical (e.g. deep brain stimulation) interventions can alleviate some of the symptoms, but lose their efficacy over time. The distinct loss of DA neurons in the SN offers the opportunity to assay neuronal cell replacement, and the clinical transplantation of fetal midbrain neuroblasts in PD patients has shown that this approach is feasible. However, there are multiple problems associated with the use of fetus-derived material, including limited availability. DA neurons derived from stem cells (SC) represent an alternative and unlimited cell source for cell replacement therapies. Currently, human pluripotent SC, such as embryonic (ES), and most recently, induced pluripotent stem cells (iPS), and multipotent (tissue-specific) adult SC are available, although the methodology for a reliable and efficient production of DA neurons necessary for biomedical applications is still underdeveloped. Here, we discuss some essentials for SC and SC-derived DA neurons to become therapeutic agents.

27. STEM CELL THERAPY FOR HAIR LOSS Recent technology for the treatment of Alopecia (Baldness) i.e. Male & female pattern hair loss & hair loss associated with various disorders. It is a revolutionary treatment for hair fall. The stem cells promote new hair growth within 3 to 4 weeks of treatment . The hair density increases by 30 to 40% by three sessions of treatment. On an average six sessions are required once in a week to 10 days along with which supplements are given. Stem cells activate the dead hair follicles and convert them into growing healthy new hair follicles.

28. BENEFITS OF ADULT STEM CELL RESEARCH Easy to obtain Potentially limitless in supply Patients can use their own stem cells for treatment and therapy Adult stem cells are politically neutral Not offensive to any major interest group nor do they generate controversy.

29. LIMITATIONS OF ADULT STEM CELL The isolation of some types of ASC, for example the isolation of neural cells from a patient's brain, would be impractical. Where a person suffers from a genetic disorder or some types of cancers, ASC isolated from that individual will retain the damaging genetic alterations underlying the disease and so be of little therapeutic value. Maintenance in culture is difficult.

30. REFERENCE www.stemcellcentre.edu.au www.stemcellchannel.com.au Hiremath; M. B. and Nerli; R. B. (2008). “Advances in Stem Cell Research”. CURRENT SCIENCE, VOL. 95, NO. 3, 10 AUGUST 2008. Bio-Medicine (2009). “Life Cell in India can store Cord Blood Stem Cells and Save Live”. Retrieved from [Online] Available http://www.bio-medicine.org/medicine-news/-u2018Life-Cell-u2019-In-India-Can-store--Cord-Blood- Stem-Cells-and-Save-Live--6033-1/ Gupta, DK et al. Stem cell therapy: Hope and scope in pediatric practice. J of Indian Assoc Pediatric Surgery. 2005:10;138-41. Biotechnology- V. Kumaresan.