Stem cell culture

Stem cell culture, their types, important

1. Stem Cells, Types and their Culture Praveen garg VITS College, Satna

2. A stem cell line is a group of stem cells that is cultured in vitro and can be propagated indefinitely. Stem cell lines are derived from either animal or human tissues and come from one of three sources: embryonic stem cells, adult stem cells or induced pluripotent stem cells. Stem cell, an undifferentiated cell that can divide to produce some offspring cells that continue as stem cells and some cells that are destined to differentiate (become specialized). Stem cells are an ongoing source of the differentiated cells that make up the tissues and organs of animals and plants.  INTRODUCTION

3. Stem cell have potential in the development of therapies for replacing defective or damaged cells resulting from a variety of disorders and injuries, such as Parkinson disease, heart disease, and diabetes . Bone marrow contains cells called hematopoietic stem cells, which generate all the cell types of the  blood and the immune system.  Hematopoietic stem cells are also found in small numbers in peripheral blood and in larger numbers in  umblical cord blood . 

4. They have unlimited self-renewal capabilities. They are non-differentiated cells with unspecialized functions. They can differentiate into specific cell types under the right conditions. 5 Types of Stem Cells by Differentiation Potential: Totipotent (or Omnipotent) Stem Cells. Pluripotent  Stem Cells. Multipotent  Stem Cells. Oligopotent  Stem Cells. Unipotent  Stem Cells. TYPES AND FEATURES OF STEM CELLS

5. They can differentiate into embryonic, as well as extra-embryonic tissues, such as chorion , yolk sac, amnion, and the allantois . The most important characteristic of a totipotent cell is that it can generate a fully-functional, living organism. Example of a totipotent cell is a fertilized egg.   Totipotent Stem Cells

6. It can self renew and differentiate into any of the three germ layers: ectoderm, endoderm, and mesoderm. These three germ layers further differentiate to form all tissues and organs. Among the natural pluripotent stem cells,  embryonic stem cells  are the best example. Another a type of “human-made” pluripotent stem cell also exists, which is the   induced pluripotent stem cell ( iPS cell).  iPS cells are tissue specific cells that can be reprogrammed to become functionally similar to embryonic stem cells. Induced pluripotent stem cells are well suited for use in cellular therapy and regenerative medicine. Pluripotent Stem Cells

7. Multipotent stem cells are a middle range type of stem cell, in that they can self renew and differentiate into a specific range of cell types. An excellent example of this cell type is the mesenchymal stem cell (MSC). Mesenchymal stem cells can differentiate into osteoblasts (a type of bone cell), myocytes (muscle cells), adipocytes (fat cells), and chondrocytes (cartilage cells). These cells types are very diverse in their characteristics. Multipotent Stem Cells

8. Oligopotent cells, are similar to the multipotent stem cells, but they become further restricted in their capacity to differentiate. While these cells can self renew and differentiate, they can only do so into closely related cell types. An excellent example of this cell type is the hematopoietic stem cell (HSC). HSCs are oligopotent stem cells derived from mesoderm that can differentiate into both myeloid and lymphoid cells. Myeloid cells include  basophils , dendritic cells, eosinophils , erythrocytes, macrophages, megakaryocytes , monocytes , neutrophils , and platelets, while lymphoid cells include B cells, T cells, and natural kills cells. Oligopotent stem Cells

9. The unipotent stem cells, which are the least potent and most limited type of stem cell. An example of this stem cell type are muscle stem cells . While muscle stem cells can self renew and differentiate, they can only do so into a single cell type. They are unidirectional in their differentiation capacity. Unipotent  Stem Cells

10. Stem cells can be originate from these cells: Embryonic stem cells Adult stem cells Induced Pluripotent stem cells Sources of stem cells

11. Around 3–5 days after a sperm fertilizes an egg, the embryo takes the form of a blastocyst or ball of cells. The blastocyst contains stem cells and will later implant in the womb. Embryonic stem cells come from a blastocyst that is 4–5 days old. The embryo ( blastocyst ), contains an outer cell mass that become part of placenta and an inner cell mass that is capable of generating all the specialized tissues that develop into the human body.  ESCs are derived from the inner cell mass of an embryo that has been fertilized  in vitro . It is not derived from eggs fertilized in a woman’s body.  This is known as pluripotent stem cells have the potential to become any cell type and are only found during the first stages of development.  These stem cells grown in vitro such as nerve, skin, intestine, liver, etc for transplantation.  1. Embryonic stem cells (ESCs)

12. Embryonic stem cell culture

13. A body contains stem cells throughout their life. The body can use these stem cells whenever it needs them. ASCs are undifferentiated, multipotent cells found in living differentiated tissues in our bodies that can renew themselves or generate new cells that can replace dead or damaged tissue.  It is also called tissue specific or somatic stem cells, adult stem cells exist throughout the body from the time an embryo develops. ASCs are present in different tissue such as the brain, bone marrow, blood and blood vessels, umbilical cord, placenta, skeletal muscles, skin, the liver, fat tissue etc. ASCs generate new cells to replace those that are lost through normal repair, disease, or injury. 2. Adult stem cells (ASCs)

14. Hematopoietic Stem Cells (Blood Stem Cells) Mesenchymal  Stem Cells. Neural Stem Cells. Epithelial Stem Cells. Skin Stem Cells. Types of Adult Stem Cells ( multipotent cells)

15. Hematopoietic stem cells (HSCs) are  multipotent , have self renewal capacity and the ability to regenerate all the different blood forming cells. hPSC culture requires growth factors, cell to cell interactions and cell to matrix adhesions. Media enriched with growth factors found in fetal bovine serum (FBS) or defined serum replacements. hPSCs are grown in aggregates, or colonies, which helps create this niche. hPSC culture systems utilize support cells such as an inactivated mouse embryonic fibroblast (MEF) feeder layer to support growth and prevent differentiation. These cells provide necessary intercellular interactions, extracellular scaffolding and factors creating a stable hPSC culture environment. Hematopoietic stem cells culture

16. MSCs exhibit the potential for differentiation into a variety of different cells/tissue lineages. (MSCs) are  multipotent stem cells found in bone marrow that are important for making and repairing skeletal tissues, such as cartilage, bone and the fat found in bone marrow. These are not to be confused with haematopoietic (blood) stem cells that are also found in bone marrow and make our blood. MSCs come from the connective tissue or stroma that surrounds the body’s organs and other tissues. MSCs has to be used to create new body tissues, such as bone, cartilage, and fat cells. They play a role in solving a wide range of health problems. Mesenchymal stem cells (MSCs)

17. Neural stem cell, largely undifferentiated, self-renewing, multipotent cells originating in the central nervous system.  Neural stem cells (NSCs) have the potential to give rise to offspring cells that grow and differentiate into neurons and glial  cells. When transplanted, NSCs are able to improve the phenotype in different transgenic models of motor neuron disease. Neural stem cells can help to treat things such as stroke, spinal cord injury, and Parkinson’s disease (a disease in which cells that contribute to control body movements progressively stop working and die).  Neural stem cell

18. Epithelial stem cells (EPSCs) are a  multipotent cell and self renewal. Epithelial stem cells are responsible for everyday regeneration of the different layers of the epidermis. These stem cells are found in the basal layer of the epidermis. Many epithelial tissues are capable of regeneration, that is, they are capable of rapidly replacing damaged and dead cells. Epithelial stem cells

19. Epithelial stem cell culture

20. Skin stem cells are multipotent adult stem cells present in the adult skin. It can self-renew and differentiate into different cell lineages of the skin.  Skin stem cells are active during skin renewal, which occurs throughout life, and in skin repair after injury. The epidermis of the skin contains layers of cells called keratinocytes . Only the basal layer, next to the dermis, contains cells that divide. Hair follicle stem cells ensure constant renewal of the hair follicles. The stem cells at the base of the skin stop proliferating and start differentiating into the cells that form the skin itself. Skin stem cells

21. Induced pluripotent stem cells are stem cells that are created in the laboratory, a happy medium between adult stem cells and embryonic stem cells.  IPSCs are created through the introduction of embryonic genes into a somatic cell (a skin cell) that cause it to revert back to a “stem cell like” state.  These cells, like ESCs are considered pluripotent discovered in 2007, this method of genetic reprogramming to create embryonic like cells, is novel and needs many more years of research before use in clinical therapies.  3. Induced pleuripotent stem cells (IPSCs)

22. The protocols include: Gelatin Coating of Culture Plates. Thawing and Seeding of Frozen Inactivated Mouse Embryonic Fibroblasts ( iMEFs ) Thawing and Seeding of Pluripotent  Stem Cells onto a Mouse Embryonic Fibroblast ( iMEF ) Feeder Layer. Replacement of Medium for Pluripotent  Stem Cell Culture.

23. Diseases Treated with Stem Cell Transplants Acute leukemia. Amegakaryocytosis or congenital thrombocytopenia. Aplastic anemia or refractory anemia. Chronic lymphocytic leukemia. Familial erythrophagocytic lymphohistiocytosis . Osteopetrosis .

24. THANK YOU