slide1
Download
Skip this Video
Download Presentation
Proteomic characterization of early-stage differentiation of mouse embryonic stem cells into neural cells induced by all trans retinoic acid in vitro

Loading in 2 Seconds...

play fullscreen
1 / 17

PowerPoint ?????? - PowerPoint PPT Presentation


  • 72 Views
  • Uploaded on

Proteomic characterization of early-stage differentiation of mouse embryonic stem cells into neural cells induced by all trans retinoic acid in vitro. Electrophoresis 2001, 22, 3067-3075. Presented by Shin jina . Origin of human pluripotent stem cells.

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about 'PowerPoint ??????' - ariane


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript
slide1

Proteomic characterization of early-stage differentiation of mouse embryonic stem cells into neural cells induced by all trans retinoic acid in vitro

Electrophoresis 2001, 22, 3067-3075

Presented by Shin jina

slide2

Origin of human pluripotent stem cells

Stem cell : undifferentiated (unspecialized) cellsthat can renew

themselves and also give rise to one or more specialized

cell types with specific functions in the body

slide5

cDNA array images of the expression pattern of genes in ES cells and differentiation cells

- 18 down regulation

- 61 up regulation

Molecular Reproduction and Development 56: 113-123 (2000)

slide6

Purpose

  • To gain more insight about the molecular mechanism of early neurogenesis and neural differentiation, we combined the advantages of ES cell in vitro differentiation system and proteomic analysis
slide7

morula

Mouse embryo genesis and time course of ES cell

differentiation

In vivo

blastocyst

ATRA treatment

ICM

-LIF

SEB

CEB

ESC

In vitro

4 days

4 days

10 days

Differentiation cells

Control(ES cell)

RA4

EB4

EB1

slide8

Differentiation of ES cells into neural cells at a high efficiency induced by ATRA in vitro

A

B

neurofilament

Neural cell from the brain of new-born rat

Differentiated cells

slide9

ATRA treatment

-LIF

SEB

CEB

ESC

In vitro

4 days

4 days

10 days

Differentiation cells

Control(ES cell)

RA4

EB4

EB1

Mouse embryo genesis and time course of ES cell

differentiation

slide10

Comparison of proteomic patterns between parent ES cells and ES-differentiared cells by silver staining 2-DE

A

B

Control (856 spots)

EB1 (703 spots)

D

C

EB4 (767 spots)

RA4 (668 spots)

slide11

Isolation of proteins associated with early-stage differentiation of ES cells into neural cells by coomassie brilliant blue R-250 staining 2-DE

B

A

Control

RA4

1-10: down regulation

11-24: up regulation

slide12

Identification of proteins associated with early-stage differentiation of ES cells into neural cells by PMF

slide13

24 protein spots

- protein match: 12 (table3)

- unidentified protein: 12

slide14

Vimentin expression alteration during ES cell differentiation into neural cells

Control ES cells

EB1

EB4

RA4

Vimentin: cytoskeleton protein,beginning marker of neurogenesis

while neurofilament proteins begin to be expressed, vimentin

expression is gradually lost

slide15

Follistatin-relative protein expression alteration during ES cells differentiation into neural cells

Control ES cells

EB4

EB1

RA4

(Silver stain)

Follistatin-relative protein: neural proliferation and survival factor

negative regulation of cellular growth, take part in triggering of ES

cells into differentiation into neural cells induced by RA

slide16

Other proteins expression alteration during ES cells differentiation into neural cells

  • Tubulin a-3/7: cytoskeleton protein, different tubulin isoforms might play different roles in neurogenesis
  • Cytokeratin 8/18: typical marker of neural cells
  • NEL-like protein: contains several EGF-like repeats, it might function as a ligand by stimulating other molecules such as EGF receptor in a neuron-specific signaling pathway
  • Unknown proteins: further investigation and which identification might shed more light about the detail of molecular mechanisms of neurogenesis
slide17

Discussion

  • Identified some important regulators in neurogenesis by comparing proteome of 4 stage cells during in vitro neurogenesis
  • The strategy provides a model to analyze effects of cytokines, growth factors or hormones on early neuronal development
  • Favorable for investigating complex mechanisms regulating neuronal differentiation
  • A further challenge will be to determine how these factors involved in the process of neuronal cell differentiation or brain development
ad