Storage Diseases and Derivation of Mutant StemCell Lines by Preimplantation Genetic Diagnosis. Gheona A ltar escu, ...
INTRUDUCTIONPreimplantation genetic diagnosis( PGD) is a great tool foravoiding the transmission for genetic dise...
Stem cells• Undifferentiated cell capable of self- renewal and differentiate into more specialized cells.
Stem cells • Self-renewal: the ability to go through numerous cycles of ...
Types of stem cells1. Embryonic stem cells, which are isolated from the inner cell mass of blastocysts2. Adult stem cell...
Lysosomes
lysosomes ClassificationOver 60 lysosomal enzymes • Primary: those thatare ...
LSD• Lysosomal storage diseases are genetic disorders in which a genetic mutation affects the activity of one or more of...
PGDPreimplantation genetictesting is a technique usedto identify genetic defectsin embryos createdthrough i...
Objective• Present the strategy and outcome of PGD for four lysosomal storage disorders ( TSD,GD,FD,HS) with the purpose...
Materiales y Métodos• Tay Sachs 4 (PGD)-5(prenatal diagnosis) Do...
• Gaucher Familia 2 : heterocigoto /mutación paterna Familia 3:mujer GS/marido mutación 84GG -50%
• Fabry :2 parejas : hombre enfermo de fabry azoospermia no obstructiva en una pareja hombres ...
IVF- estimulación ovárica, recuperación delovocito , fertilización y biopsia congelados-...
Análisis Molecular• Se extrajo el ADN de las células de sangre periférica ( parejas, niños afectados y familiares de prim...
Derivación de líneas y mantenimiento• Derivación y mantenimiento de indiferenciados Shaare Zedek (SZ) Hunter y células de...
Tabla 3
Tabla 3
Tabla 4• De los 28 embriones, se obtuvieron dos líneas de células madre embrionarias humanas (HESC).• Una de un e...
Figura 1
Figura 1 (A )Electroforesis: método delaboratorio en el que seutiliza una corrienteeléctrica controlada con lafinal...
Figura 1 (b)• Análisis cromosómico por tinción de Giemsa, llevado a cabo en metafase.• Mostró un cariotipo norma...
Figura 1 (c) • Se observaron colonias de células madre embrionarias humanas (HESC) y cue...
Discussion A GREE OR A UTHOR OPINION ...
Discussion A GREE OR A UTOR OPINION ...
Conclusions ery interestingThe article was v t he statistics ofbecau se ...
Conclusions a It is satisfying to know that PGD is ...
Concept map
MAPAPAULA
GRACIAS
Prevention of lysosomal storage diseases and derivation of1
Prevention of lysosomal storage diseases and derivation of1
Prevention of lysosomal storage diseases and derivation of1
Prevention of lysosomal storage diseases and derivation of1
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Prevention of lysosomal storage diseases and derivation of1

Published on: Mar 4, 2016
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Transcripts - Prevention of lysosomal storage diseases and derivation of1

  • 1. Storage Diseases and Derivation of Mutant StemCell Lines by Preimplantation Genetic Diagnosis. Gheona A ltar escu, Rachel Beeri, Rachel Eiges, Silvina Epsztejn-Litman, Talia Eldar -Geva, Debor ah Elstein, A r i Zimran, Ehud J.Mar galioth, Ephrat Levy-Lahad, and Paul Renbaum.Tatiana Gil FrancoPaula E MontoyaMedicine studentsMolecular Biology
  • 2. INTRUDUCTIONPreimplantation genetic diagnosis( PGD) is a great tool foravoiding the transmission for genetic diseases todescendants. This is a challenging method because haveanalyze de disorder in a single cell , and have to buildprotocols for each specific mutation.Is necessarily that give the results in a short time, that canused in testing two or more indications at once andaccuracy rates approaching 100%; to get this, useinformation of genomic DNA from family and polar bodies ,how give maternal autosomal dominant.
  • 3. Stem cells• Undifferentiated cell capable of self- renewal and differentiate into more specialized cells.
  • 4. Stem cells • Self-renewal: the ability to go through numerous cycles of cell division while maintaining the undifferentiated state. • Potency: the capacity to
  • 5. Types of stem cells1. Embryonic stem cells, which are isolated from the inner cell mass of blastocysts2. Adult stem cells, which are found in various tissues.
  • 6. Lysosomes
  • 7. lysosomes ClassificationOver 60 lysosomal enzymes • Primary: those thatare known. There is a contain enzymes solon.hydrolyse for each type of •  Secondary: besidesbiological molecule:  containing enzymes, also•Peptidases – hydrolyse digestion materials.proteins•DNAases – hydrolyse DNA•RNAases – hydrolyse RNA•Lipases – hydrolyse lipids•Phosphatases – hydrolysephosphates•Glucosidases – hydrolyseglycogen
  • 8. LSD• Lysosomal storage diseases are genetic disorders in which a genetic mutation affects the activity of one or more of the acid hydrolases. In such diseases, the normal metabolism of specific macromolecules is blocked and the macromolecules accumulate inside the lysosomes, causing severe physiological damage or deformity.
  • 9. PGDPreimplantation genetictesting is a technique usedto identify genetic defectsin embryos createdthrough in vitro fertilization(IVF) before pregnancy.Preimplantationgenetic diagnosis (PGD)refers specifically to whenone or both geneticparents has a knowngenetic abnormality and
  • 10. Objective• Present the strategy and outcome of PGD for four lysosomal storage disorders ( TSD,GD,FD,HS) with the purpose of avoiding the transmission for genetic diseases and termination of pregnancy in couples at risk of transmitting of these disorders.
  • 11. Materiales y Métodos• Tay Sachs 4 (PGD)-5(prenatal diagnosis) Double carrier
  • 12. • Gaucher Familia 2 : heterocigoto /mutación paterna Familia 3:mujer GS/marido mutación 84GG -50%
  • 13. • Fabry :2 parejas : hombre enfermo de fabry azoospermia no obstructiva en una pareja hombres serán sanos –mujeres portadoras (ligado al X) no implantar portadores• Síndrome de Hunter1 y 2 : hermanas –CVS3: mujer –análisis prenatal –interrupción –
  • 14. IVF- estimulación ovárica, recuperación delovocito , fertilización y biopsia congelados- descongelados: Valerato de estradiol oral (Estrofem 4- 8mg al día) y vaginal Utrogestan (progesterona micronizada 900 mg / día). Cuerpo polar y blastómero
  • 15. Análisis Molecular• Se extrajo el ADN de las células de sangre periférica ( parejas, niños afectados y familiares de primer grado ).• Para cada enfermedad:Marcadores polimórficos de microsatélites que rodean el gen enfermo se identificaron y los marcadores informativos usados, se crearon haplotipos para cada familia. Estos marcadores y las mutaciones familiares se usaron para el desarrollo de ensayos múltiples de una sola célula.Una reacción de PCR múltiple se utiliza .Sólo las muestras que fueron informativas para un mínimo de tres marcadores polimórficos se consideraron para el diagnóstico.
  • 16. Derivación de líneas y mantenimiento• Derivación y mantenimiento de indiferenciados Shaare Zedek (SZ) Hunter y células de Gaucher se llevaron a cabo de acuerdo con protocolos aplican rutinariamente en blastocitos diagnosticados como mutante
  • 17. Tabla 3
  • 18. Tabla 3
  • 19. Tabla 4• De los 28 embriones, se obtuvieron dos líneas de células madre embrionarias humanas (HESC).• Una de un embrión mutante Hunter hembra.• Otra de compuestos heterocigotos 84GG/N370S para la
  • 20. Figura 1
  • 21. Figura 1 (A )Electroforesis: método delaboratorio en el que seutiliza una corrienteeléctrica controlada con lafinalidad de separarbiomoleculas según sutamaño y carga eléctrica através de una matrizgelatinosa. (1937) Estas nuevas líneas presentan características típicas de células madre embrionarias humanas, que expresan un panel de marcadores no diferenciados, como NANOG, Oct4, Sox2, y REX
  • 22. Figura 1 (b)• Análisis cromosómico por tinción de Giemsa, llevado a cabo en metafase.• Mostró un cariotipo normal, 46XX humano para la línea celular Hunter y 46XY para la línea de
  • 23. Figura 1 (c) • Se observaron colonias de células madre embrionarias humanas (HESC) y cuerpos embrioides. • HESC: células que poseen la capacidad de dividirse por largos periodos, se pueden diferenciar en las células de distintos linajes. (Pluripotentenciales). • Cuerpos embrioides: son agregaciones espontáneas de HESC que se
  • 24. Discussion A GREE OR A UTHOR OPINION DISA GREE In the second couple there was no known infertility but since more than half of female carriers of Fabry K. Toyooka disease develop symptoms during life [22], they preferred medical sex election for males. Since PGD was firstG. L. Harton, M. performed in 1991, by using different strategies, the De Rycke, F.
  • 25. Discussion A GREE OR A UTOR OPINION DISA GREEP. Kozlowski, A. Both of these invasive methods are accompaniedKnippel, and R. by a small risk of abortion Stressig. due to the procedure [28]. One Gaucher stem cell line was derived from mutant A. Ribner, G. embryos caring 84GG and Altarescu, A. N370S mutations is of Zimran, and particular of interest due to D. Elstein. recent evidence of correlations between Parkinson disease and
  • 26. Conclusions ery interestingThe article was v t he statistics ofbecau se it shows us nd od for orphan aa p revention meth l uc h as lysosomaserio us diseases, s . st orage diseases It is also interestin g to note the goo prognosis that has d this therapy in the prevention of the children’s birth wit mutations. h
  • 27. Conclusions a It is satisfying to know that PGD is procedure in w hich is cared the bility and also is embryo’s sta forecasted the fu ture mutations in genes of the embryo.Despite being a techniq ue in which is risked thelives of many embryos , is a procedure whospurpose is to provide e better quality for babieborn. s
  • 28. Concept map
  • 29. MAPAPAULA
  • 30. GRACIAS

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