title: Enhanced differentiation potential of human amniotic mesenchymal stromal cells by using three-dimensional culturing.
abstract: The therapeutic potential of human amniotic mesenchymal stromal cells (hAMSCs) remains limited because of their differentiation towards mesenchymal stem cells (MSCs) following adherence. The aim of this study was to develop a three-dimensional (3-D) culture system that would permit hAMSCs to differentiate into cardiomyocyte-like cells. hAMSCs were isolated from human amnions of full-term births collected after Cesarean section. Immunocytochemistry, immunofluorescence and flow cytometry analyses were undertaken to examine hAMSC marker expression for differentiation status after adherence. Membrane currents were determined by patch clamp analysis of hAMSCs grown with or without cardiac lysates. Freshly isolated hAMSCs were positive for human embryonic stem-cell-related markers but their marker profile significantly shifted towards that of MSCs following adherence. hAMSCs cultured in the 3-D culture system in the presence of cardiac lysate expressed cardiomyocyte-specific markers, in contrast to those maintained in standard adherent cultures or those in 3-D cultures without cardiac lysate. hAMSCs cultured in 3-D with cardiac lysate displayed a cardiomyocyte-like phenotype as observed by membrane currents, including a calcium-activated potassium current, a delayed rectifier potassium current and a Ca(2+)-resistant transient outward K(+) current. Thus, although adherence limits the potential of hAMSCs to differentiate into cardiomyocyte-like cells, the 3-D culture of hAMSCs represents a more effective method of their culture for use in regenerative medicine.
authors.affiliation: Department of Cardiology, Peking Union Medical College Hospital, ShuaiFuYuan 1#, East District, Beijing, 100730, China. seeingsea@gmail.com
authors.forename: Xue
authors.initials: X
authors.lastname: Lin
authors.forename: Hao Yu
authors.initials: HY
authors.lastname: Li
authors.forename: Lian Feng
authors.initials: LF
authors.lastname: Chen
authors.forename: Bo Jiang
authors.initials: BJ
authors.lastname: Liu
authors.forename: Yian
authors.initials: Y
authors.lastname: Yao
authors.forename: Wen Ling
authors.initials: WL
authors.lastname: Zhu
mesh_terms: D000650:Amnion / Q000166:cytology*; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D055724:Electrophysiological Phenomena; D000071063:Endoglin; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009124:Muscle Proteins / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015221:Potassium Channels / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism; D055746:Stage-Specific Embryonic Antigens / Q000378:metabolism; D034421:Sus scrofa; D014020:Tissue Extracts; D020107:Troponin T / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; D003470:Culture Media; C064206:ENG protein, human; D000071063:Endoglin; C119587:HCN4 protein, human; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D009124:Muscle Proteins; D015221:Potassium Channels; D011956:Receptors, Cell Surface; D055746:Stage-Specific Embryonic Antigens; D014020:Tissue Extracts; D020107:Troponin T; C066584:stage-specific embryonic antigen-4