Morphology of hES Cells
Colony Growth
Differentiating hES Cells
Embryoid Bodies
Human ES Cell Colony Growth – Days after passaging hES colonies
| |
Undifferentiated cell colonies retain
smooth boundaries |
 |
 |
| Day 1 |
Day 2 |
|
|
 |
|
| Day 3 |
|
Differences Between Undifferentiated and Differentiating hES Cells
Undifferentiated, pluripotent hES cell colonies
|
 |
|
 |
hES cell colonies appear as single layer translucent cell disc with clearly defined borders. |
Differentiating hES cell colonies
|
 |
|
 |
Flattening and spreading out in the center of the colony. Cells around the edge of the colony pile up. |
 |
|
 |
Dendritic-like processes extend from the edge of the differentiating colony. |
 |
hES cell colony no longer exhibits a defined border. Cells along edges of the colony have begun to enlarge and spread out. |
Further Differentiation of hES Cells to Embryoid Bodies
 |
Embryoid Body formation 3 weeks after passaging hES cells on plastic without MEF feeder layer nor FGF, but in the presence of FBS. |
 |
Molecular analysis of differentiation markers of mRNA from pluripotent hES cells (H9) and Embryoid Bodies (EB).
|
| Gene |
Forward primer (5’- 3’) |
Reverse primer (5’- 3’) |
| Oct-4 |
CGACCATCTGCCGCTTTGAG |
CCCCCTGTCCCCCATTCCTA |
| Nanog |
TGCCTCACACGGAGACTGTC |
TGCTATTCTTCGGCCAGTTG |
| FLK1 |
AAGGTGACAGGAAAAGACGAACT |
TCCCCTCCATTGGCCCGCTTAAC |
| NCAM |
AGGAGACAGAAACGAAGCCA |
GGTGTTGGAAATGCTCTGGT |
| AFP |
ACTGCAATTGAGAAACCCACTGGAGATG |
CGATGCTGGAGTGGGCTTTTTGTGT |
RT-PCR primer sequences for molecular analysis of differentiation.