Stem cells Loaded with Expectations

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Stem cells Loaded with Expectations

• -
• Petri Lehenkari
• Department of Surgery
• University of Oulu
• Finland
• -

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All healing in adult tissue is facilitated by
stem cells

• Totipotent stem cell
• -embryonal cells before division into inner and
  outer cells
• Pluripotent stem cell
• -emryonal cells (inner cells)
• Multipotent
• -adult stem cells
• -resident in bone marrow and many other tissues

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Adult multipotent stem cells somatic stem
cells have been isolated from several tissues
Leonardo Da Vinci

• Bone marrow, muscle, liver, spleen, tendon, fat,
  prepatellar fat body, brain, skin, gut

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The classical idea of cell differentiation and
tissue regeneration is challenged by the modern
cell biology
Paul Berg,Stanford Univ
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Examples of self-renewal in the adult body

• Cut mucosa renewed at 2hrs -2 days
• Skin renewal at 20 days
• Heart remodeling at 5 years
• All bones rebuilt at 10 years
• Brain tissue remodelled at 20 years
• Growing evidence suggests that all this renewal
  of human body is facilitated by adult stem cells

Le Corbusier
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Cloning of differentiated cells has revealed the
multipotency of all cells

• Any somatic cell can be dedifferentiated and
  returned into stem-cell-stage
• Growth arrest and serum starvation (protein
  synthesis depletion) restores the multipontency
  of the nucleus
• Wilmut I et al (Edinburgh, UK) Nature
• 1997 Feb 27385(6619)810-3

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Implications of the adult stem cell paradigma

• Aging
• -insufficient stem cell capacity? (dementia,
  osteoporosis, malabsorption, infections)
• Disease
• -inappropriate stem cell behavior (scarring in
  spinal chord injury, artery wall calcification
  and bone formation within, cancer)

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Stem Cell Research has two traditions

• Embryonal stem cells
• 1700 embryology
• 1997 cloning
• 1998 inner cell mass culture
• 2000 combinant techniques
• 2003- therapeutic cloning, human-mouse chimera

• Adult stem cells
• 1959 bone marrow transplantation between
  identical siblings E. Donnall Thomas
• 1968 Friedensteins exsperiments bone marrow
  cells differentiate into mesenchymal tissues
• 1970-1980 Fliedner and Körbling bone marrow stem
  cells can be mobilised into peripheral
  circulation
• 1999- Friedensteinin studies are rehabilitised at
  USA
• 2001- donor cells are found in unexpected
  tissues experimental studies begin

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Stem Cell Research in Finland

• National Technology Agency Inquiry 2003
• 35 research groups focused on stem cell research
  (140 persons)
• Both embryonic and adult stem cell research
• Resourcing by Public Bodies
• TEKES, Finnish Academy, Government directly
• Academic research accentuated at the moment

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Stem Cell ethics in Finland
Misha Gordin
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Stem Cell Group Oulu is interested in Adult Stem
Cells(somatic stem cells)

• Because their use is ethically acceptable
• Bone marrow transplantation is already at
  clinical use
• Bone marow stem cells are a hot topic and new
  applicable data is produced rapidly
• Bone marrow stem cells can easily be induced into
  bone cells
• Clinical applications for bone repair are obvious

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Oulu Stem Cell Team -aim

• To culture somatic stem cells and develop
  clinical applications

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Bone Marrow Stem Cell Cultures at OUH (Oulu
University Hospital)

• Bone marrow derived mesenchymal stem cells have
  been cultured from 146 patients (age 2-98 years)
  between 2000-2004, urothelial stem cells cultured
  from 12 patients
• Stem Cell Bank of 110 donors, SOP and GC
  protocol, ethical committee approval for
  research and 5 clinical autologous
  trasplantations for congenital pseudoarthrosis
  patients
• First autologous osteoblast trasplntation back
  into patient done at 6.4.2004

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Clinical trials

• Phase 1
• -proof of safety
• Phase 2
• -proof of effectivity
• Phase 3
• -proof of clinical usability

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NF-1 (neurofibromatosis) Bacground for patient
selection

• NF-1 Skeletal manifestations
• -osteoporosis
• -congenital tibial pseudoarthrosis

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Transplantation of cells back into patients
principle of the phase 1 clinical trials
Bone marrow is harvested from iliac crest and
grown in culture
Tibial Nonunion
2 weeks of culture
Live implants
Cells in solution
Implantation of bone marrow cells into bone
defect with or without carrier
Osteoinduction occurs, healing the bone defect?
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Cells in a carrier live implants

• ALP staining, red color bone cells

With cells
Wo cells
Cells on plastic (bone cell differentiation Marked
with red color)
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Cells embedded in the collagen carrier vs cells
in solution

• 3 microbiological check ups (bacterial etc
  cultures, pcr) are performed prior to green for
  transplantation

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Cell culture facilities are located adjescent to
the operation theatre
Operation theathres
Cell culture
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Patients own bone marrow cells are multiplied in
culture

• At beginning 100.000-200.000 mesenchymal stem
  cells from 15 ml of bone marrow
• After 2 weeks 20-30.000.000 cells, both in
  solution and in collagen carriers

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Cells embedded in the collagen carrier vs cells
in solution

• Application in practise

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Cells embedded in the collagen carrier vs cells
in solution

• Application in practice

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Results?

• Aim to prove clinical safety
• Not to prevent the amputation!
• Postoperation
• -no pain
• -no acute adverse effects
• -no acute infection problems
• Control at 5-6 months after transplantation
• -no signs of infection
• -patient nro 2 benefits from transplantation
• Patient nro 1 has healed first fracture but
  develops a new fracture

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Clinical trials

• Phase 1
• -proof of safety
• Phase 2
• -proof of efficacy
• Phase 3
• -proof of clinical usability

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Acknowledgements

• Research teams in Oulu and in Turku
• Dept of Pediatric surgery
• Children and their parents!

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The end!