Authors: Guo Kai, Luo Yan, Li Shiqing, Tian Jingyi
To evaluate the feasibility of PKH26 marked human adipose derived mesenchymal stem cells (hADSCs) in vitro and intraocular.
HADSCs were cultured in vitro and marked with PKH26.Eighteen C57BL/6J mice were divided into normal control group (6 mice), labeled cell group (9 mice) and unlabeled cell group (3 mice) by using sortition randomization method, labeled cells were injected intravitreally in C57BL/6J mice as labeled cell group, and the unlabeled cells were injected intravitreally in C57BL/6J mice as unlabeled cell group.After 1 month, retinal slab was checked to contrast the results of intraocular labeling.The retina was taken out for hematoxylin-eosin staining and electron microscopy to observe the toxicity.The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.
In vitro, red fluorescent was found in the cytomembranes after hADSCs were labeled by PKH26.Retinal patch of labeled cell group showed red fluorescence of hADSCs were in front of the retina.Hematoxylin-eosin staining of retinal tissue and optic ganglion cells showed that cell degeneration and proliferation were not found in labeled cell group.The hADSCs in vivo and in vitro marked enhancement results showed that, no fluorescent was found in the unlabeled cell group, the color positive rate were both 0, while red fluorescence was found in the labeled cell group, the color positive rate were both 100%.
PKH26 can be used to mark and intraocular trace of hADSCs.Also, it has no morphology toxic reaction.