Abstract
Purpose :
Mesenchymal stem cells (MSCs) have the ability of tissue regeneration effects, such as the improvement of cardiac function, revascularization, and intestinal tissue repair, etc. MSCs are important for tissue homeostasis and wound healing. MSCs have been reported to exist in amniotic membrane (AM), which has the ability of anti-inflammatory effect and healing promoting effect, etc. To estimate whether our isolated human AM mesencymal cells (AMMCs) were involved in the effect of AM, we examined the effect of AMMCs on human corneal epithelial sheets in vitro and rabbit corneal epithelium in vivo.
Methods :
AMs were supplied by Tokyo Dental College General Hospital Amniotic Membrane Bank. AMMCs were isolated from AM using collagenase digestion, and expanded by cultivations. AMMCs phenotype was analyzed by many cell surface markers using flow cytometer. AMMCs were also induced to the osteoblasts and neural cells, and we observed cell phenotype by cell staining and RT-PCR. Human coreal limbal epithelial sheets were prepared from donor limbal tissue using the collagenase treatment, and spread on culture insert using DMEM/F12 medium containing EGF and B27 supplement for 10 days. This medium was changed to DMEM/F12 medium without supplement or the cultured supernatant with AM or AMMCs for 4 days. These sheets were compared by histochemical analysis, colony forming efficiency, and wound healing model by developed epithelial defect using tissue punch. The effect of instillation or conjunctival injection of each medium to rabbit corneal epithelial wound healing model was compared.
Results :
Some mesenchymal stem cells and neural crest origin related markers were expressed on AMMCs cell surface. After the induction of differentiation, isolated AMMCs showed similar characteristics of osteoblasts and neural cells. AMMCs supernatant maintained limbal epithelial phenotype and immature state, and promoted wound healing in limbal epithelial sheets. AMMCs supernatant tended to promote rabbit corneal epithelial wound healing.
Conclusions :
This data may suggest that the isolated AMMCs were effective for maintaining limbal epithelial phenotype and promoting the wound healing, which may partially explain the effects of AM in ocular surface construction.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.