Amnion epithelial cell isolation and characterization for clinical use.
Human amnion epithelial cells (hAECs) are a heterologous population positive for stem cell markers; they display multilineage differentiation potential, differentiating into cells of the endoderm (liver, lung epithelium), mesoderm (bone, fat), and ectoderm (neural cells). They have a low immunogenic profile and possess potent immunosuppressive properties. Hence, hAECs may be a valuable source of cells for cell therapy. This unit describes an efficient and effective method of hAEC isolation, culture, and cryopreservation that is animal product-free and in accordance with current guidelines on preparation of cells for clinical use. Cells isolated using this method were characterized after 5 passages by analysis of karyotype, cell cycle distribution, and changes in telomere length. The differentiation potential of hAECs isolated using this animal product-free method was demonstrated by differentiation into lineages of the three primary germ layers and expression of lineage-specific markers analyzed by PCR, immunocytochemistry, and histology.
Murphy
Sean
S
Department of Obstetrics and Gynaecology, Monash University, Clayton, Victoria, Australia.
Rosli
Sharina
S
Acharya
Rutu
R
Mathias
Louisa
L
Lim
Rebecca
R
Wallace
Euan
E
Jenkin
Graham
G
D000650:Amnion / Q000166:cytology / Q000502:physiology*; D000818:Animals; D002453:Cell Cycle; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D015925:Cryopreservation / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007621:Karyotyping / Q000379:methods; D051379:Mice; D016133:Polymerase Chain Reaction; D016615:Telomere / Q000648:ultrastructure
D007166:Immunosuppressive Agents