Isolation, Characterization, Cryopreservation of Human Amniotic Stem Cells and Differentiation to Osteogenic and Adipogenic Cells

Abstract

Human stem cells and progenitor cells can be used to treat cancer and replace dysfunctional cells within a tissue or organ. The objective of this study was to identify the appropriate cells type in regenerative medicine and targeted therapy. As an alternative to embryonic and bone marrow stem cells, we examined human amniotic fluid stem cells (hAFSCs), one of the potential source of multipotent stem cells isolated from both cell pellet (using single-stage method), and supernatant of human amniotic fluid. Source of isolation and unique property of the cells emphasize that these cells are one of the promising new tools in therapeutic field. Double sources for isolation and availability of the left over samples in diagnostic laboratory at the same time have less legal and ethical concerns compared with embryonic stem cell studies. Cells were isolated, cultured for 18th passage for 6 months and characterized using qPCR and flow cytometry. Cells showed good proliferative ability in culture condition. The cells successfully differentiated into the adipogenic and osteogenic lineages. Based on these findings, amniotic fluid can be considered as an appropriate and convenient source of human amniotic fluid stem cells. These cells provide potential tools for therapeutic applications in the field of regenerative medicine. To get a better understanding of crosstalk between Oct4/NANOG with osteogenesis and adipogenesis, we used network analysis based on Common Targets algorithm and Common Regulators algorithm as well as subnetwork discovery based on gene set enrichment. Network analysis highlighted the possible role of MIR 302A and MIR let-7g. We demonstrated the high expression of MIR 302A and low expression of MIR let7g in hAFSCs by qPCR.

MIAT lncRNA is overexpressed in breast cancer and its inhibition triggers senescence and G1 arrest in MCF7 cell line

“…Emerging evidence implicates that dysregulation of lncRNAs expression plays critical role in many human diseases including cancer pathogenesis . LncRNA MIAT was deemed to be contributing in several diseases such as cardiac fibrosis, and paranoid schizophrenia but there is little information about its role in cancer cells.…”

The human amniotic fluid mesenchymal stem cells therapy on, SKOV3, ovarian cancer cell line

“…The WAF1 is downstream of the wild type p53 gene and p53 effects on its promoter region thus are regulated by the cell cycle arrest activity of p21. In tumor cells, P53 gene is mutated so that the normal function of P21 gene is compromised and disturbed (Aziz et al, 2019).…”

“…This cyclin has the role of mitogen or is a stimulant of mitosis in normal cells cycle. Mitogen growth factors cause the accumulation of cyclin D1 which stimulates Raf and PI-3K in the subsequent stages and finally causes the progression of mitosis and passing from phase G1 (Aziz et al, 2019). Cyclin D1exists in natural cells at the point of transition of cell from phase G1 to phase S during inspection points for the repair of damaged DNA and prevention of accumulation of mutations.…”

“…Cyclin D1exists in natural cells at the point of transition of cell from phase G1 to phase S during inspection points for the repair of damaged DNA and prevention of accumulation of mutations. Genetic abnormalities occur in cell cycle in most cancers in phase G1 especially for cyclins (Aziz et al, 2019). The increase in the activity of cyclin D1 in ovarian cancer is probably regulated by steroids, like estrogen.…”

Application of nanomaterials in three‐dimensional stem cell culture

“…Stem cells are the “master cells” of tissues and organs of the body, which undergo differentiation to serve mainly for restoration or replacement of worn out, injured, or diseased cells of various tissues and organs , such as heart, bone marrow, brain, muscle tissues, skin, and so on, and have been adapted for their regenerative roles . There are two major types of stem cell in mammals: embryonic stem cells (ESC) which are somatic cells capable of differentiating to become part of all cell types found in tissues and organs of the body such as heart, nervous, immune system, etc, therefore, have been termed is pluripotent .…”