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

Abstract

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.

Increased proliferation and differentiation capacity of placenta-derived mesenchymal stem cells from women of median maternal age correlates with telomere shortening

“…Human adipose-derived MSCs from young (<30 years), adult (35-50 years) and aged (>60 years) donors have been shown to exhibit an age-related decrease in MSC frequency, cell colony formation, and cellular doubling [ 20 ], as well as a decrease in proliferation and differentiation potential [ 21 ]. Similar findings were reported in MSCs isolated from dental pulp [ 22 ], bone marrow [ 23 ] and human amniotic fluid-derived stem cells (hAFSCs) [ 24 ]. hAFSCs from donors divided into 20-29, 30-39 and 40-49 years age groups demonstrated an increase in population doubling time with age, and cells from donors of younger ages showed higher cellular proliferation [ 24 ].…”

“…Similar findings were reported in MSCs isolated from dental pulp [ 22 ], bone marrow [ 23 ] and human amniotic fluid-derived stem cells (hAFSCs) [ 24 ]. hAFSCs from donors divided into 20-29, 30-39 and 40-49 years age groups demonstrated an increase in population doubling time with age, and cells from donors of younger ages showed higher cellular proliferation [ 24 ]. In contrast to previous studies, our strategy of using smaller age ranges allowed us to underscore that PDMSCs derived from donors with median ages (22-25, 26-30, and 31-35 years) showed an increased capacity for cellular proliferation, cell doubling, and colony formation in that of cells from the younger (18-22 years) and older (>36 years) age groups, demonstrating that there is an increase in self-renewal properties at median ages, followed by a decrease in proliferation potential with further increases in age.…”

“…Abumaree et al reported CD105- and CD90-positive stem cells from chorionic villi of the human term placenta using flow cytometry [ 25 ]. Furthermore, Borzou et al [ 24 ] demonstrated that hAFSCs from different age groups were positive for CD73, CD90 and CD105, mesenchymal stem cell markers, and negative for CD34 and CD45, hematopoietic markers, but few, if any, studies have investigated the effect of maternal age on the expression of MSC surface antigen markers and the differential expression of these markers in PDMSCs. Alrefaei et al measured the differential expression of CD105 and CD29 via immunohistochemistry and gene expression analysis in components of umbilical cord [ 26 ] and placental [ 10 ] tissues of 5 maternal age groups; they reported a negative correlation with maternal age, with strong cytoplasmic and nuclear CD105 expression in the placental villi from 20- to 25- and 26- to 30-year-old donors and decreased expression in donors aged >31 years.…”

Achievements and Challenges in Transplantation of Mesenchymal Stem Cells in Otorhinolaryngology

“…They are non-hematopoietic cells that are plastic-adherent and spindle shape with self-renewing, migration, and differentiation properties [16]. They express mesenchymal surface markers such as CD44, CD73, CD90, and CD105, but they lack expression of hematopoietic markers such as CD34 and CD45 [17]. Various sources were mentioned for the isolation of MSCs including adipose tissue [18], bone marrow [19], and dental pulp [20].…”

Therapeutic Effect of Mesenchymal Stem Cells in Ulcerative Colitis: A Review on Achievements and Challenges

“…MSCs were primarily recorded as a group of non-hematopoietic, self-renewing, plastic-adherent and fibroblast-like stromal cells [19] that have the ability to transdifferentiate into ectodermal and endodermal cells (e.g., chondrogenic, adipogenic, and osteogenic) [20]. They express cell surface markers such as CD44, CD73, CD90, and CD105, and lack the expression of hematopoietic markers such as CD34 and CD45 [21].…”