Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantation

Abstract

Amniotic fluid as a novel source of mesenchymal stem cells for therapeutic transplantationHuman mesenchymal stem cells (MSCs) are multipotent stem cells, able to differentiate into multiple mesenchymal lineages. [1][2][3] Previously, we have shown that human fetal lung-derived MSCs enhance the engraftment of human umbilical cord blood (UCB)-derived CD34 ϩ hematopoietic cells in nonobese diabetic-severe combined immunodeficiency mice. 1 Here we show that secondtrimester amniotic fluid is an abundant source of fetal MSCs that exhibit a phenotype and multilineage differentiation potential similar to that of postnatal bone marrow (BM)-derived MSCs. We suggest that amniotic fluid is an attractive source of MSCs for cotransplantation in conjunction with UCB-derived hematopoietic stem cells.Amniotic fluid was collected transcervically from 6 secondtrimester legal terminations of pregnancy (mean gestational age, 19 weeks [range, 17-22 weeks]) according to a protocol approved by the medical ethical review board of our hospital. Amniotic fluid samples, without visible contamination with blood, were centrifuged for 10 minutes at 1283 rpm. Pellets were resuspended and cultured as described previously. 1 Adherent cells were detached with trypsin/EDTA (ethylenediaminetetraacetic acid) and phenotypically characterized by flow cytometry using fluorescein isothiocyanate-or phycoerythrin-conjugated antibodies. The adipogenic and osteogenic differentiation capacity of culture-expanded MSCs was determined as previously reported. 1 To confirm the fetal origin of cultured cells, a molecular HLA typing was performed on DNA obtained from expanded MSCs, and fetal and maternal blood cells by polymerase chain reaction/sequence-specific oligonucleotide using a reverse dot blot method. 4 MSCs were cultured from all 6 consecutive samples of second-trimester amniotic fluid. A quantity of 2 mL amniotic fluid was sufficient to culture these cells. The expansion potential of amniotic fluid-derived MSCs exceeded that of BM-derived MSCs. As a result, we were able to expand amniotic fluid MSCs to about 180 ϫ 10 6 cells within 4 weeks (3 passages). The phenotype of the culture-expanded amniotic fluid-derived cells was similar to that reported for MSCs derived from secondtrimester fetal tissues and adult BM 1,2 (Table 1). Amniotic fluidderived MSCs showed multilineage differentiation potential into fibroblasts, adipocytes, and osteocytes. Molecular HLA typing of fetal and maternal cells confirmed that the cultured cells were of fetal origin, without detectable contamination of maternal cells (Figure 1).Following allogeneic transplantation, most studies indicate that MSCs remain of host origin, 5 possibly as a result of the low frequency of these cells in stem cell grafts. The frequency of MSCs in UCB is particularly low, and most laboratories have been unable to grow MSCs from UCB. 6,7 Supplementing stem cell grafts with MSCs to promote engraftment has been proposed. Studies in mice and sheep show that engraftment can be promoted by the addition of t...

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.…”

Phenotype and ultrastructure of stem cells derived from amniotic fluid of Nitra rabbit

“…These two markers were not proposed by International Society for Cellular Therapy (ISCT; Dominici et al, 2006), however they are often used to detect MSC populations. There are many other studies of AF-MSCs, which found theses markers to be expressed in human (In't Anker et al, 2003;Tsai et al, 2004;De Coppi et al, 2007), rabbit (Steigman et al, 2009;Klein et al, 2010;DeKoninck et al, 2013;Fei et al, 2013;Kováč et al, 2016) and other species (Park et al, 2011;Chen et al, 2011;Nadri and Soleimani, 2007;Iacono et al, 2012;Choi et al, 2013). Investigators therefore use CD29 and CD44 together with some of molecular markers proposed for human MSCs by ISCT.…”

Mesenchymal Stromal Cells Derived from Human Embryonic Stem Cells, Fetal Limb and Bone Marrow Share a Common Phenotype but Are Transcriptionally and Biologically Different

“…MSCs are obtained from several adult mesenchymal tissue sources such as: bone marrow [8], adipose tissue [9], synovial membrane [10], periosteum [11] and dental pulp [12] as well as from non-adult tissue sources such as the fetal tissue [13], amniotic fluid [14] umbilical cord tissue (Wharton's jelly) [15] and umbilical cord blood [16].…”