PMC

(A) Cell collection from the amnion. (B) Cell culture from passage 1 of amniotic membranes from the initial third 20x canine fetuses (Bar: 50 μm). Cell culture from passage 1 of amniotic membranes from the middle third of canine fetuses (Bar: 50 μm); (C, D) cell culture from passage 1 of amniotic membranes from the final third of canine fetuses (Bar: 50 μm); (E) graphical representation of the growth curve profile of canine amniotic membrane (CAM) stem cells; (F) doubling time graph indicating the number of days required for doubling the number of CAM cells; (G) colony-forming unit (CFU) test of CAM cells from the three gestational stages stained by Giemsa (Bar: 100 µm).

a No differences in approximate number of proliferative cells of both cultures were reported on the 7th day. b Significantly more cells were observed in the mitosis phase of the amniotic cell culture. c The number of population doubling in the amniotic cell culture in the 21st day after the seeding has significantly increased. d The average ± SD speed of amniotic cells migration was higher than that of the adipose-derived regenerative cells (ADRCs)

Schematic overview of the procedure used to collect large volumes of term amniotic fluid using a closed catheter-based system, followed by MNC isolation and cell culture

Piece of amniotic membrane (a); immunohistochemistry for HMB-45 applied to cells cultured on slide. A large number of melanocytes can be observed (b); micrograph of cross-sectioned AM stained with HMB-45 after cell culture (c). Original magnification: (b, c) X100.