Isolationcharacterization of - PowerPoint PPT Presentation

Title: Isolationcharacterization of


1
Isolation/characterization of Mesenchymal stem
Cell from Whartons Jelly
Introduction Post-natal adult stem cells are
gaining more and more prominence because of the
ethical hassles associated with usage of
embryonic stem cells. Umbilical cord blood
derived stem cells are already being banked.
Umblical cord vessels and sorrounding matrix is
derived from the embryonic mesoderm. This study
thus aims at isolating and understanding the
properties of mesenchymal stem cells from
whartons jelly, the connective tissue matrix
surrounding the perivascular spaces of human
umbilical cord. Whartons jelly derived stem
cells could be a more primitive source of stem
cells demonstrating all mesenchymal properties
like that of bone marrow derived stem cells.
Isolation of Whartons Jelly Stem Cells
Transcriptional profiles of Whartons jelly stem
cells
ABCG2
Umbilical cord Blood from arteries and vein was
flushed out with heparin containing PBS Cord was
cut open and the peri-vascular spaces was
subjected to collagenase digestion
Ficoll-hypaque gradient centrifugation was
performed to isolate mononuclear cells Cells
cultured in DMEM-LG with 10 FBS and
antibioitics Characterization
100 bp ladder
Vimentin
Sox2
hTert
Rex1
Nanog
Oct1
Tdgf
Transcript analysis Differentiation
Cell surface analysis
Cell surface characteristics of whartons jelly
stem cells
Isotype Control
HLA-DR
CD 123
CD 117
CD 13
CD 34
CD 45
CD 31
CD 50
CD 54
SSEA4
CD 90
CD 44
CD 73
CD 95
CD 21
CD 10
Osteoblast differentiation
Alizarin red
Von Kossa staining