master of molecular and developmental stem cell biology · 2012-11-30 · master of molecular and...

Master of Molecular and

Developmental Stem Cell Biology

Professor Dr. rer. nat. Dr. h. c. Beate Brand-Saberi

Department of Anatomy and Molecular Embryology

Institute of Anatomy, Faculty of Medicine

How do complex tissues

form from a few cells?

Mesodermal

stem cells Myoblasts

Myotubes,

Satellite cells Muscle fibers

Pax3

Pax7

Myf5

CXCR4

Myf5

MyoD

Mef2

Myf5

MyoD

Mef2

Myf5

MyoD

Mef2

Pax3

Pax7

Myf5

CXCR4

Mrf4

Myogenin

Pax7 (in satellite cells)

Muscle-specific proteins

Myf5

MyoD

Mef2

Myosin heavy chain

Desmin

Development of skeletal muscles

Paraxial mesoderm

somite

Somite sclerotome

and dermomyotome

Ventrolateral

dermomyotome limb

muscles

Development of skeletal muscles

Yusuf and Brand-Saberi, 2012

Wnt 1, 3a,4

Shh Noggin Fgf8

Wnt 6

BMP2,4

Intermediate Mesoderm Paraxial Mesoderm Neural Tube Notochord

BMP4

H.J. Jacob, Bochum/C. Hornik

Local signals determine the developmental fate

of somitic derivatives

Notch/

Delta

Vizualization of gene expression at mRNA-level by in situ hybridization

(Dr. Verena Chankiewitz)

MyoD is a bHLH transcription factor conserved during evolution

Nautilus in Drosophila

MyoD in vertebrates

NCBI: Probes for Functional Genomics

Brown C. In situ hybridization with riboprobes: an overview for veterinary pathologists.

Vet Pathol. 1998 May;35(3):159-67. PMID: 9598579

Satellite cell and its activation

Mitotically quiescent small mononuclear progenitor cells

Reside in a niche between the basal lamina and the sarcolemma of

their associated mature muscle fiber

Rudnicki, M A; et al. 2009

Modulpraktika im Schwerpunkt Molekulare

Biochemie der Stammzellen

• In situ Hybridisierung zur Darstellung der Expression von Entwicklungskontrollgenen im Vertebratenembryo

• Muskelstammzellen

und Myogenese

Abteilung für Anatomie und Molekulare Embryologie

Local signals determine the developmental fate

of somitic derivatives

Poland Syndrome is described as an

absence or underdevelopment of the

chest muscle (M. pectoralis) on one

side of the body and webbing of the

fingers (cutaneous syndactyly) of the

ipsilateral hand).

CXCR4/SDF-1 axis is involved in the migration of muscle precursor cells

into the limb bud

SDF-1:

Stromal-cell derived factor 1

Alpha-chemokine

CXCR4:

G protein-coupled receptor

SDF-1 is its sole ligand

Modulpraktikum in Wintersemester 2012/13 RUB

Isolation and in vitro differentiation

of murine embryonic stem cells

Dr. B. Wang & Dr. M. Napirei

Department of Anatomy and Molecular Embryology, RUB

Zygote Blastocyte

Cell culture

inner cell mass

ES cells

trophectoderm

blastocoel

Establishment of a murine embryonic stem (ES) cell line

A B

C D

Isolation and cultivation of blastocytes on mitotically arrested

murine embryonic fibroblasts (feeder cells)

Hatching of blastocyst from the zona pellucida and subsequent fast

upgrowth of the inner cell mass

Inner cell mass ready for trypsin treatment and transfer into the

24 well plate

Establishment of ES cell lines by picking out and culturing the colonies

separately

Establishment of a murine embryonic stem (ES) cell line

2 days old EB formed in the hanging-drop culture

4 days old EB in the static suspension culture

Differentiation of ES-cells by the embryoid body (EB) method

Culture of ES cells in hanging-drops

Formation of EBs in the hanging-drops

Culture of EBs in suspension medium

A B

C D

Isolation and cultivation of blastocytes on mitotically arrested murine embryonic

fibroblasts (feeder cells)

Hatching of blastocyst from the zona pellucida and subsequent fast upgrowth of the

inner cell mass (ES cells)

Inner cell mass ready for cell singling by trypsin treatment and

transfer on fresh feeder cells

Establishment of ES cell lines by picking and culturing separate ES cell

colonies

Establishment of a murine embryonic stem (ES) cell line

EB cells spontaneously differentiate into multipotent stem cells of the three germinal layers and subsequently to

terminally differentiated body cells

Abteilung für Anatomie und

Molekulare Embryologie

Klassische

Embryologie Entwicklungsbiologie

am Tiermodell

Hühnerembryo

Zebrafisch

Maus F

oto

: E

sth

er

Stö

ckli,

Zü

rich

Transmissions-

elektronenmikroskopie

Rasterelektronen-

mikroskopie

Molekularbiologie

Konfokalmikroskopie

Methoden