Li Wang, Tingting Gu, Xin Rui, Huafeng Pan, Zhongliang Cheng, Xiaoming Xu*
Hwa Mei Hospital, University of Chinese Academy of Sciences, Ningbo 315000, Zhejiang Province, China
Objective: This study aimed to observe the morphological changes in interstitial cells of Cajal (ICC) transfected with a stem cell leukaemia gene (SCL) containing a green fluorescent protein (GFP) in vitro in a high-glucose environment.
Method: To observe the effect of the green fluorescent protein-containing SCL gene, recombinant lentivirus was transfected into the bladders of guinea pigs with diabetic cardiomyopathy (DCP) on the morphology of ICC. First, the recombinant lentivirus was constructed with the SCL gene and its titer calibrated. Then, guinea pig bladder ICC were isolated and cultured in vitro. The cells were treated with 2 mmol/L, 12 mmol/L and 22 mmol/L glucose for 24 hours and then divided into a blank control group (with 1% PBS), an empty lentivirus control group (with empty lentivirus) and a lentivirus transfection group (with lentivirus carrying the SCL gene) before being transfected for 3, 5 and 7 days. The ICC state and quantity, and the GFP expression were observed using a laser confocal microscope.
Results: The number, distribution and ultrastructure of ICC in the bladder tissue of DCP guinea pigs were observed, demonstrating that the typical morphology of bladder ICC and cell morphology damaged by high glucose concentration could be observed under microscope and revealing that specific c-kit receptors could be successfully expressed by immunofluorescence.
Conclusion: Lentivirus transfection successfully transferred the SCL gene into ICC. At 15 mmol/L glucose concentration, introducing the SCL gene gradually restored the morphologically damaged ICC. However, the SCL gene did not restore the number of ICC.
Urethral Perfusion, stem Cell leukaemia gene recombination, diabetic cardiomyopathy, guinea pig bladder, interstitial cells of cajal.