Isolation of Mesenchymal Stem Cells of Fetal or Maternal Origin from Human Placenta

Abstract

Recently we reported that second‐trimester amniotic fluid (AF) is an abundant source of fetal mesenchymal stem cells (MSCs). In this study, we analyze the origin of these MSCs and the presence of MSCs in human‐term AF. In addition, different parts of the human placenta were studied for the presence of either fetal or maternal MSCs. We compared the phenotype and growth characteristics of MSCs derived from AF and placenta. Cells from human second‐trimester (mean gestational age, 19+2 [standard deviation, ± 1+3] weeks, n = 10) and term third‐trimester (mean gestational age, 38+4 [standard deviation, ± 1] weeks, n = 10) AF, amnion, decidua basalis, and decidua parietalis were cultured in M199 medium supplemented with 10% fetal calf serum and endothelial cell growth factor. Cultured cells were immunophenotypically characterized, the adipogenic and osteogenic differentiation capacity was tested, and the growth kinetics were analyzed. The origin of fetal and maternal cells was determined by molecular human leukocyte antigen typing. We successfully isolated MSCs from second‐trimester AF, amnion, and decidua basalis as well as term amnion, decidua parietalis, and decidua basalis. In contrast, MSCs were cultured from only 2 out of 10 term AF samples. The phenotype of MSCs cultured from different fetal and maternal parts of the placenta was comparable. Maternal MSCs from second‐trimester and term decidua basalis and parietalis showed a significantly higher expansion capacity than that of MSCs from adult bone marrow (p < .05). Our results indicate that both fetal and maternal MSCs can be isolated from the human placenta. Amnion is a novel source of fetal MSCs, likely contributing to the presence of MSCs in AF. Decidua basalis and decidua parietalis are sources for maternal MSCs. The expansion potency from both fetal and maternal placenta‐derived MSCs was higher compared with adult bone marrow–derived MSCs.

Mesenchymal stem cells from amnion and amniotic fluid in the bovine

“…Our in vitro differentiation studies support the findings already reported in humans (In 't Anker et al 2004, De Coppi et al 2007) and in equine species (Lange-Consiglio et al 2012): cells isolated from the amniotic epithelial portion, as well as those obtained from the AF, show high plasticity, being able to differentiate into multiple germ layers (mesoderm and ectoderm). In particular, according to data obtained from human amniotic epithelial cells (Miki et al 2005(Miki et al , 2007a, we prove the ability of bovine amnion-derived cells to undergo astrocyte differentiation, as demonstrated by GFAP expression following neurogenic induction.…”

“…In particular, MSCs obtained from human amnion have been shown to retain immunomodulatory properties as well as to strongly inhibit T lymphocyte proliferation and to survive when transplanted in immunocompetent animals without inducing any tumorigenic effect in vivo (Avila et al 2001, Kubo et al 2001, Sankar & Muthusamy 2003, Yuge et al 2004. As amnion-derived progenitor cells, MSCs from AF (AF-MSCs) are thought to be in an intermediate stage between embryonic stem cells and lineage-restricted adult stem cells (In 't Anker et al 2004, Delo et al 2006, De Coppi et al 2007, Sessarego et al 2008, Gucciardo et al 2009), originating from several fetal tissues, including skin, digestive, respiratory, and urinary systems (Prusa & Hengstschlager 2002, You et al 2009, Da Sacco et al 2010. AF-MSCs are known to play a role in preventing rejection of the fetus and are thought to have low immunogenicity (Wang et al 2006).…”

Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential

“…After these treatments, AFCs gave rise to adipocytes and osteoblasts, when fibroblasts were the predominant cell population. In this regard, it has been reported [10,12,13] that amniotic fluid mesenchymal stem cells display a multilineage differentiation potential into fibroblasts, adipocytes and osteocytes. We suggest that the limited differentiation potential towards mesenchymal lineages that we found in our AFC cultures could be due to the cell heterogeneity with high percentage of epithelial cells.…”

“…Amniotic epithelial cells obtained from cesarean sections have been shown to express markers for neuronal, glial and progenitor cells and to differentiate into neuron-like cells in the ischemic brain of adult rats [7][8][9]. Moreover, there is evidence that amniotic fluid contains fetal mesenchymal stem cells with a multilineage differentiation potential [10][11][12][13]. In this study we extensively characterized unselected amniotic cells and tested their multilineage differentiation capacity in vitro.…”

Activity of mesenchymal stem cells in therapies for chronic skin wound healing

“…A variety of easy means of isolating and expanding these cells ex-vivo (bone marrow, 15,16 adipose tissue, 16,17 placenta, 18 peripheral blood, 19 and others) also makes MSCs useful cells for therapeutic approaches to supplementing tissue regeneration ( Table 2). Additionally, these cells have been shown to have notable immunomodulatory effects on the surrounding environment following transplantation, [29][30][31][32] and can support native cells with the secretion of a variety of pro-survival and pro-migratory cytokines and growth factors.…”