The ESCs characterized were positive for Oct-4, SSEA-1
and integrin [alpha]6, while negative for SSEA-4.
Immunoreactivity of transcription factors c-Myc, Oct3/4, c-kit, and SSEA-1 was increased in all BDL groups as compared with the control group (p<0.05-0.001) (Figure 4, 5).
Identification of the stem cells can be made by immunohistochemical staining of the markers expressed during early stages (e.g., SSEA-1) or gene products (e.g., Oct4, alkaline phosphatase, etc.) (1,15,30).
en 6,0 6,5-7,5 8,5-9,5 10,5-11,5 dpc Fragilisl + (6,25) + + + c-Kit/SCF - + (7,5) + + Stella - + (7,5) + + Blimp 1 + (6,25) + + + Vasa (MVH) - - - + (10,5) DAZL - - - + (11,5) SSEA-1
- + + + TNAP - + (7,25) + + GCNA-1 - - - + (10,5) TEX101 - - - - Mili - + en relacion a linea germinal masculina Marcador S.F.
In passages 3, 5, and 20, the positive proliferation rate and SSEA-1 expression were detected.
The round and "ES-like" shape of the cell clusters was confirmed as the cells, which have the pluripotency potential by the endogenous alkaline phosphatase staining and the immunofluorescence of SSEA-1. The cell cluster is round in shape but not as round and compact as human or mouse embryonic stem cells.
Therefore, to investigate the pluripotency of our pigSSC cell lines, we immunocytochemically analyzed them for AP-activity, and expression of SSEA-1, NANOG, and OCT-4 proteins that characterize undifferentiated stem cells (Figure 5).
These colonies also stained positive for AP-activity and expressed pluripotent markers such as OCT-4, NANOG, SSEA-1 and SOX2.
Paganelli et al., "Expression profile of the embryonic markers nanog, OCT-4, SSEA-1
, SSEA-4, and frizzled-9 receptor in human periodontal ligament mesenchymal stem cells," Journal of Cellular Physiology, vol.
On the following day, the cells were dissociated and stained with SSEA-1
After being washed three times with phosphate buffered saline (PBS) and blocked with 10% normal donkey serum, the samples were incubated overnight with primary antibodies including OCT4, NANOG, and SSEA-1
(Santa Cruz Biotechnology, Dallas, TX) at 4[degrees]C.
Phenotypically, MSCs express CD44, CD73, CD146, CD166, CD90, CD29, CD105/SH2/CD1-5, vimentin and endoglin, SSEA-1
, and SSEA-4 [14,15].