STEM CELL MIDTERM REVIEW

1. Describe how to isolate and classify stem cells.

Embryonics stem Induced pluripotent Adult stem cells

cells (ESCs) stem cells (iPSCs)

Isolation Isolated from the Generated by Isolated from various adult

inner cell mass of reprogramming tissues, including bone marrow,
blastocysts, which somatic cells to a adipose tissue, umbilical cord
are early-stage pluripotent state blood, and dental pulp, among
embryos using transcription others
factors

Origin Derived from the derived from the skin Present in differentiated tissue
early stages of the or blood cells that and organs of adults and
embryo have been children
development reprogrammed back
into an embryonic-
like pluripotent

Potency Pluripotent Pluripotent Multipotent

Function Potential in Easy to obtain and Cannot differentiated into

properties differentiated of differentiate several kind of specific cells
various cells type

Surface SSEA-1 (stage- SSEA-1, Oct-4, and Hematopoietic stem cells

marker specific embryonic Nanog (HSCs) found in bone marrow
antigen 1), Oct-4 express markers such as CD34
(octamer-binding and CD45
transcription factor
4), and Nanog Mesenchymal stem cells
(MSCs) express markers such as
CD73, CD90, and CD105

2. How do you characterize stem cells?

Property Embryonics stem Induced Adult stem cells

cells (ESCs) pluripotent stem
cells (iPSCs)

Self-renewal High capacity High capacity Limited

Pluripotency Pluripotent Pluripotent Multipotent

Undifferentiated Characterized by a Like ESCs Within their tissue

State high nucleus-to- niche
 cytoplasm ratio and
prominent nucleoli.

Marker expression Oct4, Sox2, and Oct4, Sox2, and CD34 for hematopoietic
Nanog Nanog stem cells and CD133
for neural stem cells

Capacity for Highest potential to High potential to Limited (compared to

Tissue differentiate into any differentiate into ESCs & iPSCs) to
Regeneration cell type of the body any cell type of differentiate into cell
the body types of their tissue
origin

3. What are the similarities and differences between ESCs and iPSCs?
Similarities:
Pluripotency: Both ESCs and iPSCs have the ability to differentiate into cells of all three
germ layers (endoderm, mesoderm, and ectoderm), making them pluripotent.
Self-Renewal: Both types of stem cells can self-renew indefinitely under appropriate
culture conditions, maintaining their undifferentiated state.

Differences:
ESCs iPSCs
Source of Origin Derived from the inner cell Generated by reprogramming
mass of blastocysts, which are somatic cells (e.g., skin cells) into a
early-stage embryos pluripotent state using specific
transcription factors bypassing the
need for embryos
Immunogenicity Allogeneic transplantation of Autologous transplantation of
ESCs may trigger immune iPSC-derived cells eliminates the
rejection in recipients, as they risk of immune rejection, as they
are derived from a different can be derived from the patient's
individual own cells
Genetic and Possess a unique genetic and Retain some epigenetic memory of
Epigenetic epigenetic signature reflective their tissue of origin, which can
Differences of their embryonic origin. affect their differentiation potential
and function
Developmental Represent an early Retain some characteristics of their
Stage developmental stage and may somatic cell origin, which may
exhibit higher pluripotency influence their differentiation
compared to iPSCs propensity and efficiency
4. Distinguish between Normal Stem cells and Cancer Stem Cells. What are
Stem Cells markers of Mesenchymal Stem Cells (MSCs) and cancer stem
cells?

Feature Normal stem cells Cancer stem cells

Function Tissue development & Tumor initiation & growth

repair

Differentiation Into various cell types Into various cancer cell types

Self-renewal Regulated Uncontrolled

Proliferation Under controlled conditions Often display dysregulated

proliferation

Tumor Do not form tumors Initiate & sustain tumor growth

formation

Markers Specific for cell types No single marker

Stem Cells markers of Mesenchymal Stem Cells (MSCs):

CD73 (Cluster of Differentiation 73)
CD90 (Thy-1 Cell Surface Antigen)
CD105 (Endoglin)
CD44 (Hyaluronate Receptor)
CD29 (Integrin beta-1)
CD166 (Activated Leukocyte Cell Adhesion Molecule)
Stro-1 (Stromal precursor antigen 1)
SSEA-4 (Stage-specific embryonic antigen 4)

Stem Cells markers cancer stem cells:

CD133 (Prominin-1)
CD44 (Hyaluronate Receptor)
CD24 (Heat Stable Antigen)
ALDH (Aldehyde Dehydrogenase)
CD166 (Activated Leukocyte Cell Adhesion Molecule)
EpCAM (Epithelial Cell Adhesion Molecule)
ABCG2 (ATP-binding Cassette Sub-family G Member 2)
CD34 (Hematopoietic Progenitor Cell Antigen)
Nestin
Oct4 (Octamer-binding Transcription Factor 4)

5. Describe the cell cycles and explain why cell cycle checkpoints are
necessary.
Interphase: the first 3 phases of the cell cycle together undergoing cell growth and DNA
replication(G1, S and S2):
 - Cells carrying on normal activities
- Chromosomes aren't visible (not coiled up)
- Normal cell metabolism and processes are occurring
- Occurs before mitosis. Includes phases of cell cycle 'getting ready' for mitosis
G1 (Gap 1): This is the first growth stage of cell cycle and in this phase cell increases
in size by accumulating chromosomal DNA, proteins and sufficient energy
S phase (Synthesis): Chromosomes duplicated and DNA replication proceed.
Chromosomes needs to be copied before a cell divides, so that each new cell has the
correct amount of DNA and the correct number of chromosomes
G2 phase: Cell continues growing and preparation for mitosis.

M phase: Cell growth and protein production have stopped. The cells energy is used to
division of the nucleus into 2 nuclei in one cell and make 2 daughter cells (splitting or
original cell into 2)
Prophase: The nucleolus and nuclear membrane disappears, chromatids begin to coil
more tightly.
Metaphase: chromosomes are maximally condensed, arrange in a line on equatorial
plane
Anaphase: The chromosomes separate evenly to both sides of the cell
Telophase: The chromosomes uncoil, the nuclear membrane and nucleus appear

Cytokinesis: separation of the cytoplasmic components into two daughter cells.

Checkpoint:
Cell cycle checkpoints (G1, S, G2, and M phases) are crucial regulatory mechanisms
that monitor DNA integrity, prevent errors in cell division, and coordinate cell
proliferation with cellular processes and environmental conditions. They ensure
accurate progression through the cell cycle, maintain genome stability, regulate cell
proliferation, coordinate with cellular processes, respond to environmental stress, and
contribute to developmental regulation.
→ The lack of functional cell cycle checkpoints can have profound consequences for
cellular and organismal health, including genomic instability, cancer development,
increased disease risk, altered cell fate decisions, and developmental abnormalities.

6. Why are stem cell imaging and tracking methods important in stem cell
research and applications? List these methods.
Stem cell imaging and tracking methods involve visualizing and tracking stem cells using
various techniques. It enables researchers to monitor the location, migration, and
behavior of transplanted stem cells in living organisms or in vitro settings.
Cell imaging and tracking methods are important in stem cell research and applications
because:
- Seeing how stem cells behave: track movement, differentiation, and engraftment to
understand their function and improve therapies
- Monitoring treatment: see if transplanted cells reach the target site and work as
expected
- Safety Checks: Track cells to avoid unexpected migration or uncontrolled growth
- Developing new therapies: Observe how cells interact with their environment to
design better treatments

Methods:
Fluorescence Microscopy: Stem cells can be labeled with fluorescent dyes or proteins,
allowing their visualization under a fluorescence microscope.

Magnetic Resonance Imaging (MRI): Stem cells can be labeled with contrast agents
such as superparamagnetic iron oxide nanoparticles (SPIONs), enabling their detection
via MRI.

Bioluminescence Imaging (BLI): Stem cells can be genetically modified to express

bioluminescent proteins, allowing their detection using specialized imaging systems.

Positron Emission Tomography (PET): Stem cells can be labeled with radioactive
tracers, such as fluorodeoxyglucose (FDG), for detection using PET imaging.

Single-Photon Emission Computed Tomography (SPECT): Similar to PET, stem

cells can be labeled with radioactive tracers and imaged using SPECT scanners.

Optical Coherence Tomography (OCT): This imaging technique uses light to

visualize stem cells and their behavior in tissue.

Ultrasound Imaging: Stem cells can be labeled with microbubbles or other contrast
agents for visualization using ultrasound.

Nuclear Magnetic Resonance (NMR) Spectroscopy: This technique can be used to

track labeled stem cells by detecting changes in their molecular environment.

Photoacoustic Imaging: Stem cells labeled with light-absorbing nanoparticles can be

detected using photoacoustic imaging, which converts absorbed light into acoustic
signals.

Raman Spectroscopy: This spectroscopic technique can provide molecular

information about labeled stem cells, allowing their detection and tracking.