přenosy jader j. fulka, jr. cbttn a vÚŽv praha fertilization – transformation of meiotic cells...

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Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha

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Page 1: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Přenosy jader

J. Fulka, Jr.

CBTTN a VÚŽV

Praha

Page 2: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Fertilization – transformation of meiotic cells into mitotic cells

Gametes – terminally differentiated and highly specialized

Genomes - (oocyte, sperm) transcriptionally inactive after fertilization

• One cell stage embryo is totipotent

• Fertilization represents RADICAL REPROGRAMMING OF BOTH GENOMES

Page 3: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

A) Sheep

B) Mouse

C) Human

D) Rabbit

E) Bovine

F) Pig

G) Monkey

Page 4: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Mouse embryo

Human embryo

PB

MPNFPN

FPN

MPN

5-MeC

Page 5: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Human embryo

Page 6: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

PB

PN

PN MPN

FPN

49% of human 1-cell stage embryos showed abnormal demethylation pattern

Page 7: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

2PB

FP

MP

MP

Page 8: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Wilmut, I et al. (1997)

Viable offspring derived from fetal and adult mammalian cells.

Nature 385, 810-813

Page 9: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 10: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

A

B

AB

AB

AB

AB

Page 11: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 12: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 13: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 14: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Enukleace

Page 15: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Přenos jádra

Page 16: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 17: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 18: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 19: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 20: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 21: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 22: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

What do we need??

Page 23: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Oct4 Cdx2

DAPI merge

Page 24: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Nanog

Page 25: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Oct4 DAPI

Page 26: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 27: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 28: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

DifferentiationβIII tubulin

TROMA 1

DAPI

MF20

+ EB formation

Page 29: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 30: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 31: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 32: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 33: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 34: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Problém: 300 oocytů na jednu linii/ 20 – 30 dárkyň

Page 35: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 36: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Pacient ESc linie

Page 37: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated

Producing primate embryonic stem cells bysomatic cell nuclear transfer

Nature - doi:10.1038/nature06357

…. potřebujeme vůbec přenosy jader?

Page 38: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
Page 39: Přenosy jader J. Fulka, Jr. CBTTN a VÚŽV Praha Fertilization – transformation of meiotic cells into mitotic cells  Gametes – terminally differentiated
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