|
KMID : 0894520080120030221
|
|
Development & Reproduction
2008 Volume.12 No. 3 p.221 ~ p.229
|
|
|
Stem Cell Properties of Human Umbilical Cord-derived Stem Cells after Cryopreservation
|
|
Kang Hyun-Mi
Park Se-Ah
Yoon Jin-Ah
Heo Jin-Yeong
Kim Hae-kwon
|
|
|
Abstract
|
|
|
|
For the clinical application, it is needed to keep characteristics of stem cells after storage for a long time. In the present study, we examined stem cell properties of human cord-derived stem cells (HUC) after cryopreservation. Cells were isolated from human umbilical cord and cultured in vitro. At passage 2 or 3, HUC were suspended at a concentration of 1.0×10?/㎖ in cryomedium consisting of DMSO and FBS. After freezing at －80℃ overnight, HUC were cryopreserved at －196℃ nitrogen gas. After 6 months, HUC were thawed and cultured in vitro. Assessment for the stem cell properties was made upon the morphology, population doubling time, and expression profiles of genes and various proteins. Cryopreserved HUC showed more than 70% viability and maintained fibroblast-like morphology similar to HUC before cryopreservation. Throughout the culture, they underwent average 42.8 doublings and produced 6.75×1018 cells. RT-PCR analyses showed that cryopreserved HUC expressed Oct-4, nanog, SCF, NCAM, nestin, GATA-4, BMP4, and HLA-1 genes. They did not express Brachyury and HLA-DR genes. Immunocytochemical studies showed that cryopreserved HUC reacted with antibodies against SSEA-3, -4, Thy-1, vimentin, fibronectin, HCAM, ICAM, HLA-1 proteins. They did not react with antibody against HLA-DR protein. Theses genes and proteins expression patterns of cryopresserved HUC were similar to those of HUC before cryopreservation. These results suggest that cryopreserved HUC could retain proliferative potential and they expressed various genes and proteins similar to HUC before cryopreservation. Thus, cryopreservation might be useful for HUC for future research and clinical application.
|
|
|
KEYWORD
|
|
|
Cryopreserved HUC, Mesenchymal stem cells, RT-PCR, Immunocytochemical study
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|