Term amniotic fluid: an unexploited reserve of mesenchymal stromal cells for reprogramming and potential cell therapy applications

Abstract

BackgroundMesenchymal stromal cells (MSCs) are currently being evaluated in numerous pre-clinical and clinical cell-based therapy studies. Furthermore, there is an increasing interest in exploring alternative uses of these cells in disease modelling, pharmaceutical screening, and regenerative medicine by applying reprogramming technologies. However, the limited availability of MSCs from various sources restricts their use. Term amniotic fluid has been proposed as an alternative source of MSCs. Previously, only low volumes of term fluid and its cellular constituents have been collected, and current knowledge of the MSCs derived from this fluid is limited. In this study, we collected amniotic fluid at term using a novel collection system and evaluated amniotic fluid MSC content and their characteristics, including their feasibility to undergo cellular reprogramming.MethodsAmniotic fluid was collected at term caesarean section deliveries using a closed catheter-based system. Following fluid processing, amniotic fluid was assessed for cellularity, MSC frequency, in-vitro proliferation, surface phenotype, differentiation, and gene expression characteristics. Cells were also reprogrammed to the pluripotent stem cell state and differentiated towards neural and haematopoietic lineages.ResultsThe average volume of term amniotic fluid collected was approximately 0.4 litres per donor, containing an average of 7 million viable mononuclear cells per litre, and a CFU-F content of 15 per 100,000 MNCs. Expanded CFU-F cultures showed similar surface phenotype, differentiation potential, and gene expression characteristics to MSCs isolated from traditional sources, and showed extensive expansion potential and rapid doubling times. Given the high proliferation rates of these neonatal source cells, we assessed them in a reprogramming application, where the derived induced pluripotent stem cells showed multigerm layer lineage differentiation potential.ConclusionsThe potentially large donor base from caesarean section deliveries, the high yield of term amniotic fluid MSCs obtainable, the properties of the MSCs identified, and the suitability of the cells to be reprogrammed into the pluripotent state demonstrated these cells to be a promising and plentiful resource for further evaluation in bio-banking, cell therapy, disease modelling, and regenerative medicine applications.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0582-6) contains supplementary material, which is available to authorized users.

Mesenchymal stem cells: Cell therapy and regeneration potential

“…(Batsali et al, 2017;Beeravolu et al, 2017;Im et al, 2005;Jin et al, 2013;Vishnubalaji et al, 2012) PB Up to P5 27 hr 50% (Jumi Kim, Shin, Jeon, Chung, & Chae, 2012) UCB Up to P18 40 hr 24% (Heo, Choi, Kim, & Kim, 2016;Jin et al, 2013;Pievani et al, 2014) CL Up to P22 54 hr 59% (Araujo et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017) WJ Up to P20 43 hr 80% (Batsali et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017;Trivanovic et al, 2013) CPJ > P40 16 hr 92% (Bakshi, McKee, Walker, Brown, & Chaudhry, 2018;Beeravolu et al, 2016;Beeravolu et al, 2017) PC Up to P10 51 hr 16% (Araujo et al, 2017;Beeravolu et al, 2016;Beeravolu et al, 2017;Heo et al, 2016) FT-blood Up to P20 50 hr ND (Campagnoli et al, 2001;In't Anker et al, 2004) FT-BM Up to P20 33 hr 34% (Brady et al, 2014;Campagnoli et al, 2001;K.-S. Shin et al, 2009;Q. Wang et al, 2016) FT-heart Up to P15 80 hr ND (Garikipati et al, 2018) FT-liver Up to P20 52 hr ND (Campagnoli et al, 2001;In't Anker et al, 2004) FT-pancreas Up to P30 30 hr ND (Hu et al, 2003) AF Up to P7 39 hr 15% (Moraghebi et al, 2017) Abbreviations: AF, amniotic fluid; AT, adipose tissue; BM, bone marrow; CL, cord lining; CPJ, cord placenta junction; FT, fetal tissue; MSCs, mesenchymal stromal/stem cells; ND; not determined; P, passage; PB, peripheral blood; PC, placenta; UCB, umbilical cord blood; WJ, Wharton's jelly 2 | ADULT MSCs Bone marrow (BM) MSCs were first discovered in 1976 by Friedenstein, Gorskaja, and Kulagina (1976). BM-MSCs have been investigated...…”

ISOLATION OF AMNIOTIC FLUID MESENCHYMAL STEM CELLS (AF-MSCs) OBTAINED FROM CAESAREAN SECTIONS

“…Another study showed isolation of amniotic fluid through cesarean section labor by using a catheter. 5,10 Amniotic fluid and ASC have many…”

Amniotic Fluid Stem Cells for the Treatment of Surgical Disorders in the Fetus and Neonate

“…Amniotic fluid‐derived MSCs are a plastic‐adherent population of c‐kit − cells that fulfill the minimal criteria for MSCs (e.g., CD34 − , CD45 − , CD73 + , CD90 + , and CD105 + ) as originally described for cells derived from bone marrow . By definition, these cells have restricted differentiation potential and have been shown to differentiate into the osteogenic, adipogenic, and chondrogenic lineages in vitro . However, increased plasticity of amniotic fluid‐derived MSCs (e.g., neural‐like cells) has also been demonstrated by some investigators, and others have found the expression of many of the same core pluripotency genes as undifferentiated embryonic cells, including OCT4 and SSEA4, albeit at lower levels in most cases .…”