They are unspecialized cells that renew themselves for long periods through

cell division
What are stem cells and why are they important?
Under certain physiologic or experimental conditions, they can be induced to
become cells with special functions such as the beating cells of the heart muscle or
the insulin-producing cells of the pancreas .
Totipotent: have unlimited capability. Can develop to all postembryonic tissues
and organs.
Pluripotent: stem cells capable of giving rise to most tissues of an organism.
Multipotent: stem cells that are specialized to give rise to a particular cell type.

Bunlar gelecekte de karnza kacak. Bunlar bilin. Totipotent hcrelerden tm doku ve
organlar geliebilir. Bunlar sonsuz kapasiteye sahiptir. Bu hcreler placenta ve fetal zara
dnebilir. Pluripotent hcreler dediimizde bunlar pek ok dokuya dnebilirler. Ama
placenta ve fetal zarlar yapamazsnz bu hcrelerle. Multipotent kk hcre dediimizde
kapasitesitesi daha da snrl belirli hcre tiplerine farkllanabilecek hcre demek. Snavda
sorulursa bu ayrntlara dikkat edin. Bir all tissues biri particular dieri most cell types.
During early divisions of fertilized egg, each daughter cell (blastomer) retains
totipotency (each embryonic blastomer is able to produce all differentiated cell
types)
Blastocyst; outer cells form embryonic membranes and the placenta, inner cell
mass forms the embryo ( at this stage not all the cells are still totipotent)
Embryonic Stem Cells (ES cells)
Gastrulation; as ICM develops daughter cells begin to acquire properties different
from one to another, specific regions are formed ( different components of the
developing embryo).
Most of the cells are plurypotent type. (about blastocyte) All of these inner cell mass are
called embriyonic stem cell. nner cell mass that are plurypotent cells are called embryonic
stem cells.
Ektoderm,mezoderm,endoderm (during gastrulation)

During gastrulation, totipotency of the cells of the ICM is lost, blastula is rearranged to form
ectoderm, mesoderm and endoderm
Totipotent stem cells of the blastomere multi/pluripotent cells of the germ layers
progenitor cells of the developing organs
As the cells become specialized into tissue, their potential becomes more limited, and they
finally differentiate to terminally differentiated cells
Germinal cells retain totipotentiality

During blastocyst
stage, if you isolate
the inner cell mass
of the blastocyst,
these cells are
pluripotent
embriyonic cells.
You differentation
these pluripotent
stem cells to so
many types of adult
cell in labarotary in
specific conditions.


sources of ES cells
ES cell lines
Excess embryos from IVF clinics
Therapeutic cloning

Embryonic stem cells (ESCs) lose potential and gain differentiated function
DETERMINATION
Most stem cells in adult tissues have reduced potential to produce cells of
different types (determined)
Retention of some toti/multi-potent cells in the tissues of adults, especially in the
bone marrow

embryonic stem cells
germinal stem cells/ fetal stem cell; (primordial germ cells located in the gonadal
ridge and mesenchyma of 5-9 wk-old human fetal tissue)
adult stem cells
progenitor cells/transit-amplifying cells
nuclear transplant stem cells

Adult stem cells are also called progenitor and transit-amplifying cells. We can get
embriyonic stem cells from embriyonic stem cell line.

injection of single sperm into oocyte cytoplasm


Embriyonic kk hcreleri nerden elde edebiliriz? Hazr kk hcre dizileri var yurtdndan
getirtip alabiliriz. Tp bebek kliniklerindeki fazla embriyolardan elde edebilirsiniz.
Therapeutic klonlama ile elde edebilirsiniz. Therapeutic klonlama dediimiz olayda olgun bir
metafazik yumurtay alyorsunuz. Klonlanacak olan canlnn(rnein ben) herhangi bir
somatic hcresini alyorsunuz. Yumurta baka bir bireyden.sonra somatic hcrenin
ekirdeini yumurtaya naklederiz. Uygun bir stimulusla uyarrz ama hcrenin blnmesini
salayp elde etmek. Embriyoyu blastocyte aamasna getiriyoruz. Blastocyte in i hcre
ktlesini izole ettik aldk. Ne elde ettik? Benim dna m tayon embriyonik kk hcre elde
ettik. Bu hcreleri pek ok dokuya dntrebiliriz. rnein benim kkrdamda
dejenerasyon var oraya bu hcreleri yerletirdik. Bu ilem teurapotic klonlamadr. Bu etik bir
uygulamadr. Fakat blastocyte i izole etmeden bir tayc anneye transfer edip bir canl
domasn salarsak bu reme amal klonlamadr. Etik olarak yasak olan budur. n vitro
fertilization= tp bebek
2 tip tp bebek teknii vardr. Biri intracytoplasmic sperm injection dieri ..

Embriyo develop from inner cell mass. These tropoektodermal cells can develop into
placenta and fetal membrane.



Dolly 1996
Clone from an adult sheep cell
Had only one success in 300 tries.
Dolly grew to maturity, and successfully had a lamb by natural means in 1998.
But Dolly seems to be prematurely old.
Methods of cloning
Nuclear replacement- Genetic material (nucleus from embryonic, fetal, or adult
cell) is removed and placed into an unfertilized egg or embryo, whose nucleus
has been removed




How are embryonic stem cells grown in the laboratory?


How are embryonic stem cells grown in the laboratory?
the cells of the inner cell mass proliferate and begin to crowd the
culture dish
they are removed gently and plated into several fresh culture dishes
(subculturing, passage)
after six months or more, the original 30 cells of the inner cell mass
yield millions of embryonic stem cells
embryonic stem cells that have proliferated in cell culture for six or
more months without differentiating, are pluripotent, and appear
genetically normal are referred to as an embryonic stem cell line.
What laboratory tests are used to identify embryonic stem cells?
characterization
growing and subculturing the stem cells for many months, this
ensures that the cells are capable of long-term self-renewal
inspect the cultures through a microscope to see that the cells look
healthy and remain undifferentiated
surface markers that are found only on undifferentiated cells (SSEA3,
SSEA4, Tra1-60 and Tra1-81)
the presence of a protein called Oct-4, which undifferentiated cells
typically make,
Oct-4 is a transcription factor, meaning that it helps turn genes on
and off at the right time, which is an important part of the processes
of cell differentiation and embryonic development (Oct-4 associated
proteins, Sox-2 and nanog are indicators of active self-renewal )
Alkaline phosphatase is a common marker that is rapidly
downregulated upon initial differentiation
examining the chromosomes to assess whether the chromosomes are
damaged or if the number of chromosomes has changed (It does not
detect genetic mutations in the cells) (for the ultimate clinical utility
of the cells, it is thought that maintenance of a normal karyotype is
essential)

Embriyonic stem cells have high telomerase activity.(a different slide)

Arkadalar imdi embriyonik kk hcrelerin zellikleri nemli.(bilmeniz
gerekiyor.)Embriyonik kk hcreleri izole ettik. Tekniini ksaca biliyorsunuz artk.
Labaratuvarda kltre ettik. Bunlarn gerekten embriyonik kk hcre olduunu ispat
etmemiz gerekiyor. Plurypotent olduklarn gsterecez. Genetik olarak normal olduklarn
gsterecez. Bunlar gsterirsek evet biz de laboratuvarmzda embriyonik kk hcre dizisi
elde ettik. Artk bunu aratrmalar iin teurapoutic amal olarak kullanabiliriz diyebiliriz.
Genetik olarak anormal bir hcre iimize yaramaz. Bunlar karakterize etmenin birka yolu
var. Bir kere miskroskopla bunlarn morfolojik olarak farkllamam hcreler olduklarn
grebiliriz. Uzun sre lmeyip kendilerini yenileyebiliyorlarsa , bu embriyonik kk hcre
lehinedir, bakn bu hcreler lmeyip kendilerini yenileyebiliyorlar. Bu hcreler gerekten kk
hcreler olabilir diyebiliriz. Karakterizyon iin en nemli ey yzey markerlar var. Snavda
benim sevdiim sorulardan birisidir: embriyonic kk hcreler ne eksprese ederler? SSEA lar
var. Oct-4 eksprese ederler. Peki adult hcreler Oct-4 eksprese ederler mi? Hayr etmezler.
Alkaline phosphatase da var. Hcreyi Oct-4 ile boyadk alkaline phosphatase ile boyadk
pozitif boyanma var. O zaman ne diyebiliriz? Evet bunlar embriyonic kk hcre. Son bir test
kalyor. Karyotiplerine bakyoruz. Normal karyotip ieriyorsa tm bu ilemler baarl olmu i
embriyonik kk hcre elde etmi oluyoruz. Daha sonra bu hcreleri dondurarak saklayabiliriz
eitli amalarla kullanabiliriz.