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Stem Cell in Pharmaceutical Research

Stem Cell in Pharmaceutical Research. Ferry Sandra. Stem Cell Characteristics. Definition of stem cells: stem cells are unspecialized cells that have two defining properties: Ability to self-regenerate/renew Ability to differentiate into different type of mature cells.

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Stem Cell in Pharmaceutical Research

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  1. Stem Cell in Pharmaceutical Research Ferry Sandra

  2. Stem Cell Characteristics Definition of stem cells: stem cells are unspecialized cells that have two defining properties: • Ability to self-regenerate/renew • Ability to differentiate into different type of mature cells

  3. Source, Potential and Type of Stem Cells/ Progenitor Cells

  4. Embryogenesis

  5. Embryonic Stem Cell Embryonic Stem Cells are taken from the Inner Cell Mass at the Blastocyst stage and cultured in a petri disk

  6. Adult Stem Cell Adult Stem Cells: undifferentiated cells found among differentiated cells of a specific tissue and are mostly multipotent cells. Sources: • Bone Marrow • Peripheral Blood • Umbilical Cord Blood • Placenta • Amniotic Fluid • Adipose Tissue • Menstrual Blood • Teeth • Etc.

  7. Progenitor Cells Progenitor Cells: relatively more differentiated and more committed cells to differentiate into particular cells. Types: • Endothelial Progenitor Cells • Neural Progenitor Cells • Cardiac Progenitor Cells • Etc.

  8. Bone Marrow & Peripheral Blood Stem Cells Granulocyte Colony Stimulating Factor (GCSF) or cyclophosphamide  mobilization to periphery

  9. Umbilical Cord Blood Stem Cell Cord blood is collected from this umbilical vein. It contains 10 times richer than bone marrow in the proportion of stem cells. • Advantages of UCB as a source for SC therapy among other sources: • Real abundance of births • Simple collection procedure • No serious ethical dilemmas • Lower risk of viral contamination and attrition in DNA quality • Longer telomeres  high potential to replicate • Low number of T-cells & immaturity of B-cells  lower incidence of GVHD • Multipotent stem cells • UCB contains: • HSCs • Mesenchymal Stem Cells (MSCs) • Endothelial Stem Cells (ESCs) • Other multipotential cells: Unrestricted Somatic Stem Cells (USSC), Cord-Blood derived Embryonic-like stem cells (CBEs)

  10. Endothelial Progenitor Cells (EPCs)

  11. EPCs Culture & Characteristics Culture of PBMNCs using three kind of cocktail medium, M, X, E Cocktail M Approximately 20-50% of Double (+) cells has been achieved from 7 days cultured PBMNC in several tested cocktail medium. UEA-1-Lectin-FITC DAPI 10xObj Putative morphology of EPC and EPC colony were observed~refer to published study (Bellik et al. 2005 & Hill et al. 2002) Di-I-ac-LDL Double (+) Cells demonstrating double (+) fluorescence were identified as differentiating EPCs.

  12. Separation CD34+ cell from fresh UCBMNC Fresh UCBMNC Pre MACS Post MACS 24.16 % 0.48% ► With this technique, we could isolate CD34+ cells (hematopoietic stem cell)

  13. Isolation and Culture Adipose Stem Cells From Lipoaspirates lipoaspirate CULTURE Morphology of the cultured adipose stem cells at day 28 Culture (Passage 5) in EGM-2 medium~similar w/ BM derived mesenchymal stem cells isolation Characterization of the cultured cells CD133 CD34 KDR

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