Isolation of amniotic stem cell lines with potential for therapy

Abstract

Stem cells capable of differentiating to multiple lineages may be valuable for therapy. We report the isolation of human and rodent amniotic fluid-derived stem (AFS) cells that express embryonic and adult stem cell markers. Undifferentiated AFS cells expand extensively without feeders, double in 36 h and are not tumorigenic. Lines maintained for over 250 population doublings retained long telomeres and a normal karyotype. AFS cells are broadly multipotent. Clonal human lines verified by retroviral marking were induced to differentiate into cell types representing each embryonic germ layer, including cells of adipogenic, osteogenic, myogenic, endothelial, neuronal and hepatic lineages. Examples of differentiated cells derived from human AFS cells and displaying specialized functions include neuronal lineage cells secreting the neurotransmitter L-glutamate or expressing G-protein-gated inwardly rectifying potassium channels, hepatic lineage cells producing urea, and osteogenic lineage cells forming tissue-engineered bone.

Isolation, characterization and differentiation of mesenchymal stem cells from amniotic fluid, umbilical cord blood and Wharton's jelly in the horse

“…The mechanisms involved in the POU5F1 regulation in ESC have been widely investigated, but in the adult stage (including post-natal and fetus stages) POU5F1 function has not been identified even though it has been associated with the undifferentiated pluripotent state of stem cell populations derived from various adult human tissues or organs such as bone marrow-derived multipotent adult progenitor cells. Furthermore, positive expression of mesenchymal markers such as CD90, CD105 and CD73; and negative expression of haematopoietic markers such as CD45, CD34 and CD14 have been observed (In't Aker et al 2003, Tsai et al 2004, De Coppi et al 2007. Recently, De Coppi et al (2007) showed that human and rodent AF-derived stem cells (AFMSCs) are positive for c-kit (CD117), that is also expressed in the heart (Beltrami et al 2003) and retina stem cells (Koso et al 2007), indicating that the c-kit receptor can identify a stem cell population within different organs.…”

“…This cytokine and its receptor have been involved in embryogenesis, carcinogenesis, spermatogenesis, melanogenesis and play an important role in the haematopoiesis during embryo development. AFMSCs are thought to originate from the developing fetus and be in an intermediate stage between ESCs and lineagerestricted ASCs (De Coppi et al 2007), but the exact origin of these cells is unclear. However, due to their adipogenic, osteogenic, myogenic, endothelial, neurogenic, hepatic, chondrogenic and renal differentiation potency (Tsai et al 2004, AFMSCs may be a readily available source similar to ESCs for large numbers of different cells progenitors (De Coppi et al 2007).…”

Recent Applications of Polymeric Biomaterials and Stem Cells in Tissue Engineering and Regenerative Medicine

“…AFSCs have a high proliferation rate and have a remarkable ability to differentiate into multiple cell types. 81 As a result, it appears that AFSCs have several advantages over other stem cell populations. Previous studies have also shown that human AFSCs can give rise to osteogenic lineages.…”

“…Our research group has isolated stem cell populations from this source, termed amniotic fluid-derived stem cells (AFSCs), and which express embryonic and adult stem cell markers. 81 The undifferentiated stem cells expand extensively without a feeder layer, and the population doubles every 36 h. Lines maintained for over 250 population doublings retained long telomeres and a normal karyotype. The AFSCs are broadly multipotent.…”

“…Cell types representing each embryonic germ layer, including cells of adipogenic, osteogenic, myogenic, endothelial, neuronal, and hepatic lineages, can be induced to differentiate from clonal human lines verified by retroviral marking. 81 Therefore, these cell lines may be classified as pluripotent stem cells as they fit the criteria, but no implication that they can generate every adult tissue. Unlike ES cells, AFSCs do not form tumors in vivo, which would be advantageous in clinical applications.…”