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


METHODS
Amniotic 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.


RESULTS
The 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.


CONCLUSIONS
The 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.
['Moraghebi|Roksana|R|', 'Kirkeby|Agnete|A|', 'Chaves|Patricia|P|', 'Rönn|Roger E|RE|', 'Sitnicka|Ewa|E|', 'Parmar|Malin|M|', 'Larsson|Marcus|M|', 'Herbst|Andreas|A|', 'Woods|Niels-Bjarne|NB|']
D050156:Adipogenesis D000653:Amniotic Fluid D002448:Cell Adhesion D002454:Cell Differentiation D002460:Cell Line D019070:Cell Lineage D049109:Cell Proliferation D002469:Cell Separation D064987:Cell- and Tissue-Based Therapy D065150:Cellular Reprogramming D004847:Epithelial Cells D005260:Female D005347:Fibroblasts D006410:Hematopoiesis D006801:Humans D007231:Infant, Newborn D059630:Mesenchymal Stem Cells D009474:Neurons D010012:Osteogenesis D039904:Pluripotent Stem Cells D011247:Pregnancy
Term amniotic fluid Mesenchymal stromal cells Cell-based therapy Regenerative medicine Cellular reprogramming Caesarean section deliveries Pluripotency Biobanking