The Effect of Age and Type of Media on Growth Kinetics of Human Amniotic Fluid Stem Cells.

Abstract

Aim: This study compared growth kinetics of human amniotic fluid stem cells (hAFSCs) in different maternal age groups and two different media of AmnioMAX and Dulbecco's modified Eagle's medium (DMEM). Materials and Methods: Three milliliters of amniotic fluid (AF) was provided from 16 pregnant women who were referred for amniocentesis from 16 to 18 weeks of gestation. Mothers were divided to 20-29 (n = 5), 30-39 (n = 5) and 40-49 (n = 6) years old age groups. AF was immediately centrifuged and the cell pellet was cultured. Cells were characterized morphologically, by flow cytometry and for osteogenic and adipogenic inductions. Population doubling time (PDT) and growth kinetics were determined. AFSCs cultured in AmnioMAX were compared in various age groups. A comparison of growth kinetics of AFSCs cultured in AmnioMAX and DMEM from 40 to 49 years old pregnant women was undertaken. Results: AFSCs were adherent to culture flasks and were spindle shape, and positive for osteogenic and adipogenic inductions and for expression of CD73, CD90 and CD105 markers, but negative for CD34 and CD45. PDT among 20-29, 30-39, and 40-49 years old women was 30.9, 38.3, and 43.9 hours, respectively showing a higher cell proliferation in younger ages. When comparing AmnioMAX and DMEM, PDT was 53 and 96.9 hours, respectively denoting to a higher cell proliferation in AmnioMAX. Conclusions: Higher proliferation and plasticity of hAFSCs were noted in AmnioMAX and in younger mothers' samples. These findings can be added to the literature and open a new avenue in regenerative medicine, when hAFSCs are targeted for cell therapy purposes.