Osteoblastic differentiation potential of human amniotic fluid-derived mesenchymal stem cells in different culture conditions

•We found that there are abundant of mesenchymal stem cells (MSCs) population exist in human amniotic fluid (hAF), which was confirmed by morphology observation, the analyzing of cell proliferation, the expression of specific mesenchymal stem cells markers, and negative expression of endothelial stem cells, amniotic stem cells, hematopoietic stem cells specific markers.•We found that isolated human amniotic fluid-derived mesenchymal stem cells (hAF-MSCs) can be induced and be differentiated into osteoblastic-like cells, which can function as bone forming cells. The osteogenic differentiation potential of hAF-MSCs was confirmed by analyzing the morphology, levels of osteoblastic specific gene and protein expression, alkaline phosphatase (ALP) activity, and extracellular matrix (ECM) calcium deposition.•We found the ability of gelatin scaffold that can promote differentiation capacity of hAF-MSCs in the osteogenic differentiation process. The upregulation of osteogenic differentiation potential of hAF-MSCs combine with gelatin scaffold compare to monolayer culture was confirmed by higher levels of osteoblastic specific genes expression, higher ALP activity, higher osteoblastic specific protein expression in immunofluorescent, and higher deposition of ECM calcium. Osteoporosis is a bone degenerative disease characterized by a decrease in bone strength and an alteration in the osseous micro-architecture causing an increase in the risk of fractures. These diseases usually happen in post-menopausal women and elderly men. The most common treatment involves anti-resorptive agent drugs. However, the inhibition of bone resorption alone is not adequate for recovery in patients at the severe stage of osteoporosis who already have a fracture. Therefore, the combination of utilizing osteoblast micro mimetic scaffold in cultivation with the stimulation of osteoblastic differentiations to regain bone formation is a treatment strategy of considerable interest. The aims of this current study are to investigate the osteoblastic differentiation potential of mesenchymal stem cells derived from human amniotic fluid and to compare the monolayer culture and scaffold culture conditions. The results showed the morphology of cells in human amniotic fluid as f-type, which is a typical cell shape of mesenchymal stem cells. In addition, the proliferation rate of cells in human amniotic fluid reached the highest peak after 14 days of culturing. After which time, the growth rate slowly d