Download A novel source for Mesenchymal stem cells

A novel source for Mesenchymal stem cells
Jayanti Tokas1, Deepika Gupta1, Divya Pasrija1, Rubina Begum1, Shalini Jain2 and Hariom
Yadav2
1Department
2NIDDK,
of Biotechnology, JMIT, Radaur, Haryana, India
National Institutes of Health, Bethesda, MD 20892, USA
Corresponding: [email protected]
FETAL STEM CELLS

Sources





Specific fetal tissues
Fetal circulation, placenta,
Amniotic fluid
Umbilical cord
Potentiality varies from pluripotent to multipotent.
FETAL STEM CELLS



Fetal stem cells

Hemopoietic stem cells (HSC)

Mesenchymal stem cells (MSC)
First-trimester blood

richer source of HSC and MSC

Greater proliferative capacity

wider differentiation ability
Can be clonally derived.
AMNIOTIC FLUID

Clear, yellowish liquid, surrounds the fetus during
pregnancy

Continuously inhaled, exhaled and urinated by the
baby.

It is 98% water and electrolytes, proteins, peptides,
carbohydrates, lipids and hormones.
AMNIOTIC FLUID

Protects the developing baby against blows to the
mother's abdomen

Allows easier fetal movement

Promotes muscular/skeletal development

Protect the fetus from heat loss.

Plays a significant defensive role
Amniotic
fluid
ROLE IN FETAL DEVELOPMENT

Contains nutritional components such as proteins,
glucose, triglyceride and cholesterol

proteins and peptides possess potent bioactivity

Growth factors in AF are transported throughout
fetal body

Helps cellular movement, organs development,
cellular growth and proliferation.
AMNIOTIC FLUID STEM CELLS

Potentiality varies from pluripotent to multipotent

Second and third trimester amniotic fluid is major source

Allow scientists to sidestep the
destroying embryos for research

Can be used for tissue repair and engineering organs

Mesenchymal stem cells quantity is high in amniotic fluid
controversy
over
HETEROGENEITY OF AMNIOTIC FLUID


Variation in

Shape

Size

Nuclear/ cytoplasmic ratio

Cytoplasmic characteristics

Cell surface properties

Biochemical properties
Some cells express markers of glial and neuronal
stem cells.
AMNIOTIC FLUID :NEW SOURCE FOR
NEURAL STEM CELLS

Amniotic fluid cells when incubated with neurogenic
medium express

CD133

Nestin

Neurofilament

CNPase

p75

Neurotrophin receptor
(the brain-derived neurotrophic factor and
neurotrophin-3)
Human amniotic fluid contains cells that express markers for neuronal
stem and progenitor cells, which harbour the potential to differentiate
into neurogenic cells
MESENCHYMAL STEM CELLS

Morphology
 Small
cell body with a few cell processes
 Large
and round nucleus with a prominent
nucleolus
A
small amount of

Golgi apparatus

Rough endoplasmic reticulum

Mitochondria

Polyribosomes .
MESENCHYMAL STEM CELLS

Detection


Surface antigens
Differentiation capacity

Multipotent- Can differentiate into
 Osteoblasts
 Adipocytes
 chondrocytes
 myocytes
 neuron-like cells.
Differentiation capacity decreases with the age of
the donor and the time in culture
MESENCHYMAL STEM CELLS AND ITS
DIFFERENTIATION CAPACITY
ISOLATION AND CULTURE

Sterile needle is inserted through the abdominal wall into the
amniotic sac

Small amount of amniotic fluid is withdrawn through needle

Amniotic fluid contains fetal cells which are separated from
amniotic fluid


Cultured
Tests are performed on cultured cells

Chemical analysis


DNA analysis
Chromosomal Analysis
MESENCHYMAL STEM CELLS : FIRST
TRIMESTER
In undifferentiated state, fetal blood, liver and bone marrow
MSCs express
 CD452

CD291

CD441
 CD142

SH21
 CD682

SH31
 CD342
 Vwf2
 HLA-DR2

SH41

prolyl-4-hydroxylase
 Laminin

Actin
 Vimentin
 Fibronectin
MESENCHYMAL STEM CELLS : FIRST TRIMESTER


MSCs differentiate into

Adipocytes

Osteocytes

Chondrocytes.
Provide novel targets for in utero cellular
and gene therapy.
MESENCHYMAL STEM CELLS : SECOND TRIMESTER

MSCs exhibit a phenotype and multilineage
differentiation potential similar to postnatal bone
marrow (BM) – derived MSCs .

Multipotent (CD29)

Pluripotent (Oct-4)
MESENCHYMAL STEM CELLS : THIRD TRIMESTER

Express Oct-4.

Samples can be collected in larger amount than the second
trimester, with a much lower risk of uterine contamination.

High renewal capacity and therapeutic applications.

Share same properties and differentiation capabilities as human
embryonic stem cells .

Differ from embryonic stem cells in promising ways :
1.
Do not produce teratomas when transplanted into animals.
2.
Low antigenicity is an advantage for cell transplantation or cell replacement
therapy.
PRESERVATION

Vitrification

Amniotic stem cell banks
VITRIFICATION
AMNIOTIC STEM CELL BANKS

Biocell center, European biotechnology company is
the first firm to harvest and preserve amniotic stem
cells

Biocell Center captures the stem cells from amniotic
fluid if the family requests and pays for the
preservation of the stem cells

Amniotic fluid withdrawn during amniocentesis is
sent to the Biocell Center laboratory. The stem cells
are frozen in liquid nitrogen and preserved in the
company's state-of-the-art cryo-bank.
“H.R. 1892—National Amniotic and Placental
Stem Cell Bank of 2007
The bill shows the importance of amniotic stem cells bank
which would

Establish a National Amniotic and Placental Stem Cell Bank for
the purpose of obtaining, storing, and making available for
research and treatment human stem cells derived from
amniotic fluid or placenta .

Will maintain a collection of at least 100,000 samples of stem
cells in order to ensure genetic diversity

Will obtain stem cells only if informed consent is provided
REGENERATIVE MEDICINE

As regenerative medicine amniotic fluid stem cells
has many applications .

Cultured MSCs have been transplanted in children
with osteogenesis imperfecta (OI), a disease
causing bone fractures and fragility.

Reduced bone fractures and increased bone
density were reported to be found when MSCs were
engrafted into the defective bone
MYOCARDIAL INFARCTION
MSCs can be engrafted at the damaged
site(s), attenuate pathologic remodeling of
the heart tissue and reduce scar size,
leading to improved post-MI cardiac
function
ETHICAL ASPECTS
Amniotic fluid solve a lot of problems

It's possible to catch amniotic stem cells without
destroying embryos

They are not necessary to choose an alternative
between donor or autologuous use
NEXT STEPS TO DO

For the future it is of great importance to obtain more insights into the
spectrum of cells contained in human amniotic fluid.

Using RT-PCR the expression of a wide variety of genes, known as
markers for stem cells, progenitor cells and differentiated cell types,
should be analysed.

Amniotic fluid contains Oct-4 positive cells, a major issue for the
future is to investigate the differentiation potential of the Oct-4
positive amniotic fluid cells

It is essential to think about strategies to isolate/enrich Oct-4 positive
cells out of amniotic fluid.
“It is an ocean full of
pearls and we need to
search these pearls only
for our benefit”.