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Daniela	
  Sanges,	
  PhD	
  
endoderm	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  	
  mesoderm	
  	
  	
  	
  	
  	
  	
  	
...
Regenera9on	
  poten9al	
  
Evolu9onary	
  scale	
  
Planaria	
  
Salamander	
  
Mammalian	
  
n  Planaria	
  have	
  an	
  amazing	
  ability	
  to	
  regenerate	
  parts	
  of	
  
themselves	
  
n  Planaria	
  reg...
n  Freshwater	
  flatworms	
  (phylum:	
  Platyhelminthes)	
  
n  Live	
  in	
  freshwater	
  under	
  leaves	
  and	
  s...
n  20-­‐30%	
  of	
  all	
  the	
  cells	
  in	
  a	
  planaria	
  are	
  
Neoblasts.	
  
	
  
n  Neoblasts	
  divide	
 ...
Three types of planaria
with their neoblasts
labeled with BrdU
BrdU	
  (bromodeoxyuridine)	
  is	
  	
  
subs9tuted	
  for...
To	
  set	
  up	
  an	
  experiment	
  of	
  
regenera9on	
  with	
  
Planaria	
  in	
  the	
  classroom	
  
n  The	
  type	
  of	
  water	
  is	
  very	
  important!	
  Fresh	
  water	
  from	
  a	
  
pond,	
  stream	
  or	
  lak...
Each	
  team	
  will	
  receive	
  3	
  planaria:	
  
	
  
n  Experimental	
  Planaria	
  #1	
  
	
   	
  Every	
  team	
...
How	
  will	
  the	
  Planaria	
  regenerate?	
  
This	
  step	
  is	
  important	
  to	
  learn	
  to	
  formulate	
  an	...
Report	
  the	
  informa9on	
  and	
  the	
  results	
  DAILY	
  
§ A_er	
  you	
  cut	
  the	
  planaria,	
  how	
  does	
  the	
  mobility	
  of	
  the	
  tail	
  
fragments	
  compare	...
§ Did	
  all	
  the	
  tail	
  fragments	
  regenerate	
  photoreceptors?	
  If	
  not,	
  
which	
  fragments	
  did	
  ...
Days	
  
Head	
   Middle	
   Tail	
  
Did	
  all	
  the	
  tail	
  fragments	
  regenerate	
  photoreceptors	
  at	
  the	...
Regenera9on	
  poten9al	
  
Evolu9onary	
  scale	
  
Planaria	
  
Salamander	
  
Mammalian	
  
High	
   Moderate	
   Low	
  
 
	
  
n  LOST	
  OR	
  ALTERED	
  GENETIC	
  PROGRAM	
  
	
  
	
  
	
  
	
  
n  SMALLEST	
  STEM	
  CELL	
  PULLS	
  
	...
What is a stem cell?

a single cell that can…

replicate itself, or…

differentiate into many cell types
Differen9a9on	
  
De-­‐differen9a9on	
  
A	
   B	
   Trans-­‐differen9a9on	
  
Pluripotent	
   Mul9potent	
  
embryonic	
  stem	
  cell	
   soma9c	
  cell	
  
pluripotent	
  genes	
  
cell	
  fusion	
  
reprogramming	
  
embryonic	
  stem	
  cell	
   soma9c	
  cell	
  
pluripotent	
  genes	
  
cell	
  fusion	
  
reprogramming	
  
WNT	
  
• Re9ni9s	
  Pigmentosa	
  
• Usher	
  Syndrome	
  
• Amaurosis	
  de	
  Leber	
  
• Glaucoma	
  
• Diabe9s	
  re9nopathy	...
Sanges	
  et	
  al.,	
  Cell	
  reports,	
  2013	
  
d0	
   d2-­‐4	
   d30	
  
Damage-­‐dependent	
  
Cell	
  fusion	
  
Wnt-­‐mediated	
  
reprogramming	
  	
  
Regenera9on	
...
§ General	
  discussion	
  on	
  9ssue	
  regenera9on	
  
§ Planaria	
  regenera9on	
  experiment	
  in	
  the	
  classr...
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
AmgenTeach Webinar 2 - Stem Cell Biology
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AmgenTeach Webinar 2 - Stem Cell Biology

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A presention by Dr Daniela Sanges during AMGENTEACH webinar 24 february 2015

Published in: Education
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AmgenTeach Webinar 2 - Stem Cell Biology

  1. 1. Daniela  Sanges,  PhD  
  2. 2. endoderm                                      mesoderm                                      ectoderm   lung                    pancreas                    heart              red  blood                      skin                      neuron  
  3. 3. Regenera9on  poten9al   Evolu9onary  scale   Planaria   Salamander   Mammalian  
  4. 4. n  Planaria  have  an  amazing  ability  to  regenerate  parts  of   themselves   n  Planaria  regenera9on  is  a  great  model  for  demonstra9ng  stem   cell  func9on  and  complexity  of  9ssue  regenera9on   n  They  can  grow  easily  in  the  lab  
  5. 5. n  Freshwater  flatworms  (phylum:  Platyhelminthes)   n  Live  in  freshwater  under  leaves  and  stones   n  Avoid  light  (“nega9ve  phototaxis”)   n  Free  living  –  not  parasites   n  Carnivores  –  eat  decaying  meat,  insects  and  larvae  (egg   yolks  or  liver  in  the  lab)   n  1-­‐20  mm  in  size   n  Long  lived   n  Use  sexual  or  asexual  reproduc9on  
  6. 6. n  20-­‐30%  of  all  the  cells  in  a  planaria  are   Neoblasts.     n  Neoblasts  divide  by  mitosis  and  can  make   more  of  themselves.    They  are  the  only   dividing  cells  in  planaria.     n  When  a  planaria  is  wounded,  neoblasts   migrate  to  the  site  and  begin  dividing     n  Neoblasts  can  become  ANY  cell  the   planaria  needs—nerve  cells,  flame  cells,   reproduc9ve  cells…   Ann. Rev. Cell. Dev. Biol. 20:725-757, 2004
  7. 7. Three types of planaria with their neoblasts labeled with BrdU BrdU  (bromodeoxyuridine)  is     subs9tuted  for  thymine  during  DNA  replica9on.   Only  dividing  cells—in  this     case  the  neoblasts—will  show     detec9ble  amounts  of  BrdU.   A.  Phagocata  sp.   B.  Girardia  dorotocephala   C.  Schmidtea  mediterranea    Dugesia  sp.  
  8. 8. To  set  up  an  experiment  of   regenera9on  with   Planaria  in  the  classroom  
  9. 9. n  The  type  of  water  is  very  important!  Fresh  water  from  a   pond,  stream  or  lake  is  best;  spring  water  is  adequate     n  Keep  cultures  in  a  cool,  dark  place     n  Feed  planaria  at  least  once  a  week     n  Change  the  water  twice  a  week,  especially  a_er  feeding     n  A_er  cu`ng  them,  the  cultures  should  not  be  fed  
  10. 10. Each  team  will  receive  3  planaria:     n  Experimental  Planaria  #1      Every  team  will  cut  this  into  half,  separa9ng        the  anterior  from  the  posterior  halves.     n  Experimental  Planaria  #2      This  will  be  cut  in  a  manner  determined  by  your  team.     n  Control  Planaria      This  will  not  be  cut.   Used  with  permission  from  UC  Museum  of  Paleontology  
  11. 11. How  will  the  Planaria  regenerate?   This  step  is  important  to  learn  to  formulate  an  HIPOTHESIS  
  12. 12. Report  the  informa9on  and  the  results  DAILY  
  13. 13. § A_er  you  cut  the  planaria,  how  does  the  mobility  of  the  tail   fragments  compare  with  the  mobility  of  the  head  fragments?   Do  they  move  the  same  or  differently?  If  they  move   differently,  why  do  you  think  that  is?   § Do  you  think  the  head  fragments  prefer  light  or  shade?   How  could  you  test  this?      
  14. 14. § Did  all  the  tail  fragments  regenerate  photoreceptors?  If  not,   which  fragments  did  not?     § Did  all  the  tail  fragments  regenerate  photoreceptors  at  the  same   rate?  If  not,  which  were  slower  and  which  were  faster?     § Did  you  no9ce  a  change  in  mobility  in  the  tail  fragments  over   9me?  What  about  the  head  fragments?     § As  the  days  went  by,  what  did  you  no9ce  about  the  color  of  the   regenera9ng  9ssue?    
  15. 15. Days   Head   Middle   Tail   Did  all  the  tail  fragments  regenerate  photoreceptors  at  the  same  rate?   If  not,  which  were  slower  and  which  were  faster?      
  16. 16. Regenera9on  poten9al   Evolu9onary  scale   Planaria   Salamander   Mammalian  
  17. 17. High   Moderate   Low  
  18. 18.     n  LOST  OR  ALTERED  GENETIC  PROGRAM           n  SMALLEST  STEM  CELL  PULLS       turning  on  the     right  genes  at  the     right  9me     in  the  cells  that  come  from  the   dividing  stem  cells  (neoblast).  
  19. 19. What is a stem cell? a single cell that can… replicate itself, or… differentiate into many cell types
  20. 20. Differen9a9on   De-­‐differen9a9on   A   B   Trans-­‐differen9a9on  
  21. 21. Pluripotent   Mul9potent  
  22. 22. embryonic  stem  cell   soma9c  cell   pluripotent  genes   cell  fusion   reprogramming  
  23. 23. embryonic  stem  cell   soma9c  cell   pluripotent  genes   cell  fusion   reprogramming   WNT  
  24. 24. • Re9ni9s  Pigmentosa   • Usher  Syndrome   • Amaurosis  de  Leber   • Glaucoma   • Diabe9s  re9nopathy   • Macular  Degenera9on   • Re9nal  Detachment   RETINAL  DEGENERATIONS  
  25. 25. Sanges  et  al.,  Cell  reports,  2013  
  26. 26. d0   d2-­‐4   d30   Damage-­‐dependent   Cell  fusion   Wnt-­‐mediated   reprogramming     Regenera9on       Sanges  et  al.,  Cell  reports,  2013  
  27. 27. § General  discussion  on  9ssue  regenera9on   § Planaria  regenera9on  experiment  in  the  classroom  (Wednesday)      Predic9on  exercise      Prac9cal  and  theore9cal  ques9on  on  the  experiment   § Mammalian  Stem  Cells  and  Reprogramming  (Thursday)   § Adult  stem  cells.  Summary  of  the  course    

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