J Vet Sci.  2006 Dec;7(4):343-348. 10.4142/jvs.2006.7.4.343.

In vitro neuronal and osteogenic differentiation of mesenchymal stem cells from human umbilical cord blood

• Park KS
• Lee YS
• Kang KS

• KMID: 1089481
• DOI: http://doi.org/10.4142/jvs.2006.7.4.343

Abstract

Mesenchymal stem cells (MSCs) have the capabilities for self-renewal and differentiation into cells with the phenotypes of bone, cartilage, neurons and fat cells. These features of MSCs have attracted the attention of investigators for using MSCs for cell-based therapies to treat several human diseases. Because bone marrowderived cells, which are a main source of MSCs, are not always acceptable due to a significant drop in their cell number and proliferative/differentiation capacity with age, human umbilical cord blood (UCB) cells are good substitutes for BMCs due to the immaturity of newborn cells. Although the isolation of hematopoietic stem cells from UCB has been well established, the isolation and characterization of MSCs from UCB still need to be established and evaluated. In this study, we isolated and characterized MSCs. UCB-derived mononuclear cells, which gave rise to adherent cells, exhibited either an osteoclast or a mesenchymal-like phenotype. The attached cells with mesenchymal phenotypes displayed fibroblast-like morphologies, and they expressed mesenchymal-related antigens (SH2 and vimentin) and periodic acid Schiff activity. Also, UCB-derived MSCs were able to transdifferentiate into bone and 2 types of neuronal cells, in vitro. Therefore, it is suggested that the MSCs from UCB might be a good alternative to bone marrow cells for transplantation or cell therapy.

Keyword

differentiation; human; mesenchymal cell; stem cell; umbilical cord blood

MeSH Terms

Acid Phosphatase/metabolism
Bone and Bones/*cytology
Cell Differentiation/*physiology
Cell Separation/methods
Fetal Blood/*cytology
Humans
Immunohistochemistry
Immunophenotyping
Infant, Newborn
Mesenchymal Stem Cells/*cytology
Microscopy, Phase-Contrast
Neurons/*cytology
Periodic Acid-Schiff Reaction

Figure

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  Fig. 1 Initially adherent mesenchymal-like cells grew as spindle-shaped or stellate-shaped cells that developed into multi-polar fibroblastoid cells. They gradually reached confluency at about 30 days. A; Primary culture day 14. B & C; Primary culture day 21. D; Primary culture day 30. B, C and D shows cell clusters. A & D: ×100, B & C: ×200.

• 

  Fig. 2 Cytochemical analysis shows that the mesenchymal-like cells were positive for PAS, but they were negative for AP activity. The immunophenotyping of these cells showed positivity for mesenchym-related antigen SH2. A; SH-2 (Endoglin) staining with the mesemchymal-like cells. B; Phase contrast image, C; PAS staining with mesenchymal-like cells. D; Negative control (counterstained with hematoxylin). ×200.

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  Fig. 3 Osteoclast-like cells were positive for AP activity, but they were negative for PAS. Osteoclast-related antigen CD51/61 (the vitronectin receptor) was also expressed. A; There was AP staining with the osteoclast-like cells. B; Negative control (counterstained with hematoxylin). C; Immunofluorescence assay for CD51/61 with osteoclast like cells. D; Phase contrast. ×200.

• 

  Fig. 4 Expression of the bone phenotype after exposure of UCB mesenchymal-like cells to differentiation stimuli. A, B & C; UCB MSCs' was morphologically changed (day 14), Phase contrast, ×200. D, E & F; UCB MSCs' osteogenesis as confirmed by calcium accumulation, Von-Kossa staning, ×200.

• 

  Fig. 5 Pattern of the expression of neural specific antigens in UCB mssenchymal-like cells. A; Stained with NSE. B; Phase contrast image. C; Stained with GFAP. D; Phase contrast. (×200).

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