Evaluation indexes for the viability of umbilical cord-derived mesenchymal stem cells before transplantation

doi:10.3969/j.issn.2095-4344.2013.32.016

Abstract

Abstract:

BACKGROUND: Umbilical cord-derived mesenchymal stem cells are gaining more attention in clinical treatments. Cell viability prior to transplantation has a direct impact on clinical prognosis. Despite trypan blue staining is a widely performed procedure to assess the viability of umbilical cord-derived mesenchymal stem cells, it cannot reflect the functional capacity of those cells accurately because of some subjective factors.
OBJECTIVE: To explore sensitive and accurate assay for the functions of umbilical cord-derived mesenchymal stem cells.
METHODS: Human umbilical cord-derived mesenchymal stem cells were isolated and cultured in vitro. Cultured umbilical cord-derived mesenchymal stem cells were preserved in 0.9% saline for 0, 2, 4 and 6 hours at 4 ℃. Various methods (trypan blue staining, AnnexinV-PI, terminal deoxynucleotidyl transferase dutp nick end labeling, cell counting kit-8, live-dead assay, cell adherent assay) were used to determine the viability of post-storage umbilical cord-derived mesenchymal stem cells, and the results were compared with colony-forming efficiency, a measure of cell function.
RESULTS AND CONCLUSION: Human umbilical cord-derived mesenchymal stem cells cultured in vitro showed a spindle shape and attached growth, the third-generation umbilical cord-derived mesenchymal stem cells were positive for CD29, CD44, CD105, and negative for CD 34 and CD 45. Umbilical cord-derived mesenchymal stem cells incubated in the adipogenic and osteogenic medium were both positive. Cell viability measured with trypan blue correlated moderately with colony-forming efficiency, while the percentage of viable cells measured with other methods correlated better with colony-forming efficiency, among which adherent assay was the most obvious. It is proved that cell adherent assay-measured viability is the most accurate indicator.

Key words: stem cells, umbilical cord/umbilical blood stem cells, umbilical cord-derived mesenchymal stem cells, cell transplantation, in vitro, cell preservation, preservation effect, cell viability, cell function, colony formation, stem cell photographs-containing paper

CLC Number:

R394.2

Lei Xin, Chen Yan, Zhang Jian-lin, Cui Lei, Niu Yu-hu, Niu Bo. Evaluation indexes for the viability of umbilical cord-derived mesenchymal stem cells before transplantation[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(32): 5847-5854.

Figures/Tables 9

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