Impact of cryopreservation on CM-Dil labeled human umbilical cord mesenchymal stem cells
10.3760/cma.j.issn.1673-4181.2014.04.006
- VernacularTitle:冻存对CM-Dil标记脐带间充质干细胞的影响
- Author:
Yan CHEN
;
Jianjun WANG
;
Huaifang LI
- Publication Type:Journal Article
- Keywords:
Human umbilical cord mesenchymal stem cells;
Chloromethyl-benzamidodialkylcarbocyanine;
Cryopreservation;
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- From:
International Journal of Biomedical Engineering
2014;37(4):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the feasibility of labeling human umbilical cord mesenchymal stem cells (hUCMSCs) with chloromethyl-benzamidodialkylcarbocyanine (CM-Dil) and the influence of cryopreservation on the CM-Dil labeled hUCMSCs.Methods Using tissue explant to culture hUCMSCs,the morphology and growth situation of the cells were observed,and cell phenotype were identified by flow cytometry.The cells of the 3rd generation were labeled with CM-Dil.The cell morphology,growth situation and fluorescent markers were observed under fluorescent microscope.The labeled cells were cryopreserved and recovered,and the growth of cells and its fluorescent labeling conditions were observed after recovery.Results The morphology of hUCMSCs was similar to fibroblasts,spindleshaped and polygonal.The cells of the 3rd generation of hUCMSCs strongly expressed CD44,CD29,CD90 and CD73,but did not express CD34 and CD45.The mark rate of CM-Dil to label hUCMSCs reached as high as 90％ and the dye was mainly in the cell membrane.The labeled cells are in good condition and the cells after cryopreservation and recovery still have strong fluorescence expression,also the cell growth is in good condition.Conclusions Using CM-Dil to label and track hUCMSCs is simpler than other methods.Cryopreservation does not have negative influence on CM-Dil-labeled hUCMSCs.
