Stem cells Hessah Alshammari. Stem cells are distinguished from other cell types by two important characteristics. They are unspecialized cells capable.

Stem cells

Hessah Alshammari

bull Stem cells are distinguished from other cell types by two important characteristics

bull They are unspecialized cells capable of renewing themselves through cell division sometimes after long periods of inactivity

bull Under certain conditions they can be induced to become tissue- or organ-specific cells with special functions eg in bone marrow stem cells regularly divide to repair and replace worn out or damaged tissues

Stem cells types

bull Embryonic stem cells

bull Adult stem cells

bull Cancer stem cells

Embryonic stem cells

bull Embryonic stem cells derived from embryo in typically four or five days old which called the blastocyst

bull The blastocyst includes three structures the trophoblast which is the layer of cells that surrounds the blastocoel a hollow cavity inside the blastocyst and the inner cell mass which is a group of cells at one end of the blastocoel that develop into the embryo proper

bull An adult stem cell is an undifferentiated cell found among differentiated cells in a tissue or organ that can renew itself and can differentiate to yield some or all of the major specialized cell types of the tissue or organ

bull The primary roles of adult stem cells in a living organism are to maintain and repair the tissue

bull In fact adult hematopoietic or blood-forming stem cells from bone marrow have been used in transplants for 40 years

Adult stem cells

Similarities and differences between embryonic and adult stem cells

Embryonic stem cells adult stem cells

Differentiation ability

Pluripotent stem cellscan become all cell types of the body

multipotent stem cellslimited to differentiating into different cell types of their tissue

culture grown easily in culture Isolation and expansion challenging

Rejection after transplantation

Do not cause rejection cause rejection

Tumour formation after injection

tumour genesis is not tumour genesis

Colony forming form embryoid bodys MSC form colony forming units NSC form neurispheres

embryoid bodys

colony forming units

Neurispheres

Adult stem cells in human body

bull Neural stem cellsbull Heamatopeotic stem cellsbullMesenchymal stem cellsbullAdipose stem cellsbullSkin stem cellsbullLiver stem cellsbullMultipotent adult precursor cells MAPC bullMammary stem cells

Mesenchymal stem cell

bullSomatic progenitors able to self-renew and produce mesenchymal derivatives such as osteroblast adipocytes and chondrocytes

bullCD29 amp CD105 is MSCs marker

Mesenchymal stem cell isolation

Isolation of bone marrow

Lysis blood cells using buffer and culture

MSC

Biomedical applications

1- Pharmacological screening

2- Orthopaedic tissue repair (bone cartilage tendon)

3- Non-orthopaedic application (heartspinal cord)

4- Gene delivery

Pharmacological screening

Research into bone metabolism amp drug discovery such as osteoprosis (caused by decreased in bone formation and increase fatty marrow)

Misregulation in MSC differentiation Therefore identify components regulate MSC differentiation is leading to drug discovery

Orthopaedic tissue repair (bone cartilage tendon)

The Effect of Implants Loaded with Autologous Mesenchymal Stem Cells on the Healing of Canine Segmental Bone Defects SCOTT P BRUDER MD PHD BALTIMORE KARL H KRAUS DVM NORTH GRAFTON

MASSACHUSETTS VICTOR M GOLDBERG MD and SUDHA KADIYALA PHD BALTIMORE MARYLAND

Investigation performed at Osiris Therapeutics Baltimore and Tufts University School of Veterinary Medicine North Grafton

Radiographs of a segmental defect that had been treated with a ceramic cylinder that

had not been loaded with MSCs(A) and loaded with

MSC (B)

Non-orthopaedic application (heartspinal cord)

Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction

Luciano C AmadoAnastasios P SaliarisKarl H Schuleri and Joshua M Hare

MSC cardiomyoplasty augments development of new myocardium (Left) At 8 weeks after MI the subendocardial rim is thicker in the MSC group (arrows) Hematoxylineosin (HampE) stain of the subendocardial rim demonstrates cardiomyocytes in both control and MSC (times100 magnification)

Bar graph depicting enhanced thickness of subendocardial viable tissue with MSCs ( P lt 005 vs nontreated) after 8 weeks

Adipose stem cells

Adipose-derived stem cells was reported first time in 2002 by Hedrick et al

CD45 CD105 CD90 and CD29 is marker of adipose-derived stem cells So it is closely to MSC

Human Adipose Tissue Is a Source of Multipotent Stem Cells Patricia A Zuk Min Zhu Peter Ashjian Daniel A De Ugarte Jerry I Huang Hiroshi Mizuno Zeni C Alfonso John K Fraser Prosper Benhaim and Marc H Hedrick Departments of Surgery and Orthopedics Regenerative Bioengineering and Repair Laboratory UCLA School of Medicine Los Angeles California 90095 and Department of Medicine and the Jonsson Comprehensive Cancer Center Division of Hematology and Oncology UCLA School of Medicine Los Angeles California 90095 Submitted February 25 2002 Revised June 21 2002 Accepted August 23 2002Monitoring Editor Martin Raff

Isolation of adipose stem cells

Lipoaspirate

collagenease digest

washdensity gradient centrifugation

plating

Morphology and multilineage differentiation potential of ASCs

Primary ASCs exhibited a

fibroblast-like morphology as observed under a phase contrast

microscope

chondrogenic differentiation was verified byimmunofluorescence staining for the presence of collagen type II deposition

osteogenic differentiation was revealed

by ALP staining

adipogenic differentiatio

n was confirmed by positive Oil

Red O staining

Adipose stem cells application

bullPlastic amp cosmetic application eg breast reconstruction

bull As a source of non-adipogenic cell types eg Bone cartilage repair applicationsbullAs a vector for gene delivery or cell therapy

Adipose stem cells applications

Adipose-derived adult stromal cells heal critical-size mouse calvarial defects

Catherine M Cowan1Yun-Ying Shi1 Oliver O Aalami1Yu-Fen Chou2 Carina Mari3 Romy Thomas1 Natalina Quarto1 Christopher H Contag4 Benjamin Wu2 amp Michael T Longaker11The Department of Surgery Stanford University School of Medicine Stanford University 257 Campus Drive Stanford California 94305 USA

Neural stem cells in brain

Neural stem cells in the brain-the lining of the lateral ventricles-the dentate gyrus of the hippocampus

neural stem cells markers NestinSox1 Sox2 and Sox9

Neural stem cells

bull Precursors for neuronal and glial lineages

Isolation and culture of Neural stem cells

Brain digestiondetermine the

location of NSC in brain

Tissue disaggregation

by using enzyme eg accumax

Culture NSCIn serum-free medium with

EGF and bFGF

NSC form neurosphere in culture and has Sox1 amp Sox2 marker

Neural stem cells application

bull Transplantation of NSC to repair damaged tissue (trauma strok) neurodegeneration (Alzheimer) and delivery of genes

Neurosci Res 2005 Jul52(3)243-9Intravenous administration of human neural stem cells induces functional recovery in Huntingtons disease rat modelLee ST Chu K Park JE Lee K Kang L Kim SU Kim MDepartment of Neurology Clinical Research Institute Seoul National University Hospital 28 Yongon-Dong Chongro-Gu Seoul 110-744 South Korea

bull Fetal human NSC bullChemically induced lesion bullCell injectionbullAnalysis at 9 week

Phenotype is significantly improved

Limitations of neural stem cells application

bull Ethics of collection

bull Availability of healthy tissue

bull Viability of isolated cells

bull immunocompatibility

Skin stem cells

Stem cells have been found in three regions of the epidermis

bull the bulge region of the follicle

bull The sebaceous gland

bull The interfollicular epidermis

Skin stem cells properties

bull Have a high colony-forming ability in vitro

bull Long-term proliferative capabilitybull Strong adhsion to extrcellular matrixbull Small cells with a high nuclear to cytoplasmic ratio bull Divide when skin is injuredbull Express keratin 19 keratin 15 and CD34

Skin stem cells isolation

Culture conditionbull On irradiated feeder layer or ECMbull In the media containing of FCSbullPresences of FGF

Skin stem cells applications

bull Burns and skin damage

bull Doctors from the University of Sydney and Concord Hospital have formed the Sydney Burns Foundation in 2010

bull Under laboratory conditions scientists use the patients own skin stem cells and grow the top layer of the skin the epidermis

bull Professor Maitz treated farmer John Heffernan who suffered burns to 80 per cent of his body in 2006 Theyve had to replace his skin from where ever he got burnt

bull They did multiple surgeries and some areas wouldnt heal up so theyd have to go back and put in a bit of a graft on that While it will close the wound it has no elasticity it cannot sweat it cannot regulate temperature

bull Mr Heffernan has returned to farming but every day is a challenge as his skin is so fragile and easily damaged

bull The treatment for burns survivors has made great advances but still has a long way to go The hope is that one day theyll be able to grow a full three dimensional skin which can perform all the vital functions of healthy skin

bull Genetic conditions ( epidermolysis bullosa)

Mutation in laminin 5 gene (basement membrane component)

Epidermolysis Bullosa

Extensive blistering of the skin generating infected lesions

Correction of junctional epidermolysis bullosa by transplantation of genetically modified epidermal stem cells Michele De Luca et al 2006 University of Modena and Reggio Emilia Italy

bull Genetic conditions ( epidermolysis bullosa)

Experimental strategybullBiopsy from palm skinbullIsolation of stem cellsbullGenetic modification of mutated LAM5bullControl for restored LAM5 production in vitrobullTransplantation of multiple grafts (each ~55cm2)

They observed complete epidermal regeneration on both legs at day 8 and a normal-looking epidermis was maintained throughout the 1-year follow-up (Fig 3a

• Stem cells Hessah Alshammari
• Slide 2
• Stem cells types
• Embryonic stem cells
• Adult stem cells
• Similarities and differences between embryonic and adult stem cells
• Slide 7
• Adult stem cells in human body
• Mesenchymal stem cell
• Mesenchymal stem cell isolation
• Biomedical applications
• Pharmacological screening
• Orthopaedic tissue repair (bone cartilage tendon)
• Non-orthopaedic application (heartspinal cord)
• Adipose stem cells
• Isolation of adipose stem cells
• Morphology and multilineage differentiation potential of ASCs
• Adipose stem cells application
• Adipose stem cells applications
• Neural stem cells in brain
• Neural stem cells
• Isolation and culture of Neural stem cells
• Neural stem cells application
• Limitations of neural stem cells application
• Skin stem cells
• Skin stem cells properties
• Skin stem cells isolation
• Skin stem cells applications
• Slide 29
• Slide 30
• Slide 31