Amniotic Fluid Transplantation Alleviates Hematopoietic Deficits in Experimental Rat Aplastic Anemia

BACKGROUND Hematopoietic stem cell (HSC) transplantation is the most effective therapy for hematopoietic impairment. However, maintenance and self-renewal of HSCs in culture is still a central focus of HSC research. It is known that amniotic fluid contains a heterologous population of stem cells (AFSCs) and nutrients as well as various types of growth factors. We hypothesize that AFSCs may be expanded in vitro in pure amniotic fluid. MATERIAL AND METHODS Amniotic fluid with transparent appearance was harvested at embryo age of 13.5-15.5 days in rats and was placed in a Cell Culture CO₂ incubator. The cell number in the amniotic fluid was counted before and after culture of amniotic fluid. Then, the effect of amniotic fluid transplantation on 5-fluorouracil combined with busulfan induced-rat aplastic anemia was investigated. RESULTS We found that after a short time (about 30 min) culture, the number of AFSCs expanded more than 100-fold. Flow cytometry showed that there was a population of cells expressing hematopoietic markers CD45 and CD34 in addition to a higher proportional of mesenchymal stem cells (MSCs) markers CD29 and CD90. Transplantation of the expanded AFSCs possessed significant therapeutic effect in toxic chemicals induced-aplastic anemia rats, manifested by decreasing animal mortality and alleviating the reduction of the 3 lineages of hematopoietic cells in blood. CONCLUSIONS Our observation for the first time demonstrates that amniotic fluid is an excellent medium for stem cells to maintain "stemness" proliferation and provides novel evidence to support the potential use of in vitro-expanded AFSCs for the treatment of hematopoietic disorders.