Isolation, amplification and marker observation of Sca-1 positive cardiac stem cells in mice
LIU Ping-ping
LI Wen-jing
CHANG Fen
ZHAO Jing
JING Qing-chuan
Abstract：Objective To isolate and amplify the Sca-1 positive cardiac stem cells from C57BL/6 mice in vitro and to observe its markers .Methods The myocardial tissues from C 57 BL/6 mice were selected , and we separated the Sca-1 pos-itive cells by the magnetic cell sorting and amplified its passage .We observed the morphology changes in the culture process.The flow cytometry was used to test the Sca-1 +purity of the primary and fifth generation of myocardial cells .RT-PCR was used to detect the expression of multiple factors (MYH6, cTnT and GATA4) in the isolated Sca-1 positive cells and negative cells .Results The primary cultured Sca-1 positive cells were round , smaller than blood cells;the sub-cul-tured were starlike , and the pseudopodia were long , and after the fifth generation , the morphology of cells showed little difference.The Sca-1 +positive rates of primary and the fifth generation were 81%±10.4% and 90%±5%, separately (P>0.05).The relative expression of MYH6, cTnT and GATA4 in Sca-1 +cells was 0.5 ±0.21, 0.72 ±0.04 and 0.72 ±0.05, separately;while the expression in the Sca-1 -cells was 0.2 ±0.02, 0.86 ±0.06 and 0.71 ±0.05.Significant difference was found in the expression of MYH6 and cTnT (all P<0.05).Conclusion Sca-1 positive cardiac stem cells can be isolated successfully and sub-cultured stablely.Furthermore, Sca-1 +cells show high expression of MYH6.
Keywords：cardiac stem cellsmyosin heavy chain 6troponin Tmyocardial transcription factor 4magnetic cell sorting
Publication Date：2015-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 14-16 )