Stem cell in pharmaceutical research
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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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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


Source, Potential and Type of Stem Cells/ Progenitor Cells


Embryogenesis


Embryonic Stem Cell

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


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.


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.


Bone Marrow & Peripheral Blood Stem Cells

Granulocyte Colony Stimulating Factor (GCSF) or cyclophosphamide  mobilization to periphery


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)


Endothelial Progenitor Cells (EPCs)


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.


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)


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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