Differentiation of umbilical cord blood-derived multilineage progenitor cells into respiratory epithelial cells

• Published in: Cytotherapy (Oxford, England) England), 2006, Vol.8 (5), p.480-487
• Main Authors: Berger, M.J., Adams, S.D., Tigges, B.M., Sprague, S.L., Wang, X-J., Collins, D.P., McKenna, D.H.
• Format: Article
• Language: English
• Subjects: Cell Differentiation - physiology; Cells, Cultured; endodermal differentiation; Epithelial Cells - physiology; Epithelial Cells - ultrastructure; Fetal Blood - cytology; Fetal Blood - physiology; Humans; Multipotent Stem Cells - physiology; Multipotent Stem Cells - ultrastructure; Pulmonary Alveoli - physiology; Pulmonary Alveoli - ultrastructure; respiratory epithelium; stem cells; umbilical cord blood
• ISSN: 1465-3249; 1477-2566
• DOI: 10.1080/14653240600941549

Umbilical cord blood (UCB) has been examined for the presence of stem cells capable of differentiating into cell types of all three embryonic layers (i.e. endo-, ecto- and mesoderm). The few groups reporting success have typically confirmed endodermal potential using hepatic differentiation. We report differentiation of human UCB-derived multipotent stem cells, termed multilineage progenitor cells (MLPC), into respiratory epithelial cells (i.e. type II alveolar cells). Using a cell separation medium (PrepaCyte-MLPC; BioE Inc.) and plastic adherence, MLPC were isolated from four of 16 UCB units (American Red Cross) and expanded. Cultures were grown to 80% confluence in mesenchymal stromal cell growth medium (MSCGM; Cambrex BioScience) prior to addition of small airway growth medium (SAGM; Cambrex BioScience), an airway maintenance medium. Following a 3–8-day culture, cells were characterized by light microscopy, transmission electron microscopy, immunofluorescence and reverse transcriptase (RT)-PCR. MLPC were successfully differentiated into type II alveolar cells (four of four mixed lines; two of two clonal lines). Differentiated cells were characterized by epithelioid morphology with lamellar bodies. Both immunofluorescence and RT-PCR confirmed the presence of surfactant protein C, a protein highly specific for type II cells. MLPC were isolated, expanded and then differentiated into respiratory epithelial cells using an off-the-shelf medium designed for maintenance of fully differentiated respiratory epithelial cells. To the best of our knowledge, this is the first time human non-embryonic multipotent stem cells have been differentiated into type II alveolar cells. Further studies to evaluate the possibilities for both research and therapeutic applications are necessary.