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Isolation and characterization of canine umbilical cord blood-derived mesenchymal stem cells.

Seo MS, Jeong YH, Park JR, Park SB, Rho KH, Kim HS, Yu KR, Lee SH, Jung JW, Lee YS, Kang KS - J. Vet. Sci. (2009)

Bottom Line: After neuronal differentiation, the cMSCs expressed the neuronal markers Nestin, GFAP, Tuj-1, microtubule-associated protein 2, NF160.With osteogenic differentiation, the cMSCs presented osteoblastic differentiation genes by RT-PCR.This finding also suggests that cMSCs might have the ability to differentiate multipotentially.

View Article: PubMed Central - PubMed

Affiliation: Adult Stem Cell Research Center, Department of Veterinary Public Health, College of Veterinery Medicine, Seoul National University, Seoul 151-742, Korea.

ABSTRACT
Human umbilical cord blood-derived mesenchymal stem cells (MSCs) are known to possess the potential for multiple differentiations abilities in vitro and in vivo. In canine system, studying stem cell therapy is important, but so far, stem cells from canine were not identified and characterized. In this study, we successfully isolated and characterized MSCs from the canine umbilical cord and its fetal blood. Canine MSCs (cMSCs) were grown in medium containing low glucose DMEM with 20% FBS. The cMSCs have stem cells expression patterns which are concerned with MSCs surface markers by fluorescence- activated cell sorter analysis. The cMSCs had multipotent abilities. In the neuronal differentiation study, the cMSCs expressed the neuronal markers glial fibrillary acidic protein (GFAP), neuronal class III beta tubulin (Tuj-1), neurofilament M (NF160) in the basal culture media. After neuronal differentiation, the cMSCs expressed the neuronal markers Nestin, GFAP, Tuj-1, microtubule-associated protein 2, NF160. In the osteogenic & chondrogenic differentiation studies, cMSCs were stained with alizarin red and toluidine blue staining, respectively. With osteogenic differentiation, the cMSCs presented osteoblastic differentiation genes by RT-PCR. This finding also suggests that cMSCs might have the ability to differentiate multipotentially. It was concluded that isolated MSCs from canine cord blood have multipotential differentiation abilities. Therefore, it is suggested that cMSCs may represent a be a good model system for stem cell biology and could be useful as a therapeutic modality for canine incurable or intractable diseases, including spinal cord injuries in future regenerative medicine studies.

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Immunostaining of undifferentiated and neuronal differentiated cUCB-MSCs. cUCB-MSCs were immunostained with glial fibrillary acidic protein (GFAP), microtubule-associated protein 2 (MAP2), neuronal class III β tubulin (Tuj-1), Nestin and neurofilament M (NF160). Negative control was confirmed with Alexa 488 (green) and Alexa 594 (red). A: The cells were cultured with basal cultured media. B: The cells were cultured with neuronal differentiation media. C-H: Comparing to basal culture condition (undifferentiation) with neuronal differentiation condition. C, E and G: Undifferentiation; D, F and H: Neuronal differentiation. Nestin, Tuj-1 and NF160 were green. GFAP and MAP2 were red. Scale bars = 50 µm.
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Figure 2: Immunostaining of undifferentiated and neuronal differentiated cUCB-MSCs. cUCB-MSCs were immunostained with glial fibrillary acidic protein (GFAP), microtubule-associated protein 2 (MAP2), neuronal class III β tubulin (Tuj-1), Nestin and neurofilament M (NF160). Negative control was confirmed with Alexa 488 (green) and Alexa 594 (red). A: The cells were cultured with basal cultured media. B: The cells were cultured with neuronal differentiation media. C-H: Comparing to basal culture condition (undifferentiation) with neuronal differentiation condition. C, E and G: Undifferentiation; D, F and H: Neuronal differentiation. Nestin, Tuj-1 and NF160 were green. GFAP and MAP2 were red. Scale bars = 50 µm.

Mentions: Neuronal differentiation was examined according to the neuronal induction method. The cUCB-MSCs showed basically neuronal associated protein markers in the basal culture status. In the undifferentiated condition, the cUCB-MSCs slightly expressed GFAP, Tuj-1, and NF160 neuronal cell protein markers. However, the cUCB-MSCs did not express about Nestin and MAP2 (Fig. 2A). When inducted with neuronal differentiation media, the cUCB-MSCs showed positive expression patterns for Nestin, GFAP, Tuj-1, MAP2 and NF160 (Fig. 2B). Compared to the basal culture condition, the cUCB-MSCs had positive for Nestin, MAP2 with neuronal induction, but were negative prior to differentiation. These data showed that cUCB-MSCs had the ability to be inducted into glial and neuron cells under differentiation conditions (Figs. 2A-H).


Isolation and characterization of canine umbilical cord blood-derived mesenchymal stem cells.

Seo MS, Jeong YH, Park JR, Park SB, Rho KH, Kim HS, Yu KR, Lee SH, Jung JW, Lee YS, Kang KS - J. Vet. Sci. (2009)

Immunostaining of undifferentiated and neuronal differentiated cUCB-MSCs. cUCB-MSCs were immunostained with glial fibrillary acidic protein (GFAP), microtubule-associated protein 2 (MAP2), neuronal class III β tubulin (Tuj-1), Nestin and neurofilament M (NF160). Negative control was confirmed with Alexa 488 (green) and Alexa 594 (red). A: The cells were cultured with basal cultured media. B: The cells were cultured with neuronal differentiation media. C-H: Comparing to basal culture condition (undifferentiation) with neuronal differentiation condition. C, E and G: Undifferentiation; D, F and H: Neuronal differentiation. Nestin, Tuj-1 and NF160 were green. GFAP and MAP2 were red. Scale bars = 50 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2801133&req=5

Figure 2: Immunostaining of undifferentiated and neuronal differentiated cUCB-MSCs. cUCB-MSCs were immunostained with glial fibrillary acidic protein (GFAP), microtubule-associated protein 2 (MAP2), neuronal class III β tubulin (Tuj-1), Nestin and neurofilament M (NF160). Negative control was confirmed with Alexa 488 (green) and Alexa 594 (red). A: The cells were cultured with basal cultured media. B: The cells were cultured with neuronal differentiation media. C-H: Comparing to basal culture condition (undifferentiation) with neuronal differentiation condition. C, E and G: Undifferentiation; D, F and H: Neuronal differentiation. Nestin, Tuj-1 and NF160 were green. GFAP and MAP2 were red. Scale bars = 50 µm.
Mentions: Neuronal differentiation was examined according to the neuronal induction method. The cUCB-MSCs showed basically neuronal associated protein markers in the basal culture status. In the undifferentiated condition, the cUCB-MSCs slightly expressed GFAP, Tuj-1, and NF160 neuronal cell protein markers. However, the cUCB-MSCs did not express about Nestin and MAP2 (Fig. 2A). When inducted with neuronal differentiation media, the cUCB-MSCs showed positive expression patterns for Nestin, GFAP, Tuj-1, MAP2 and NF160 (Fig. 2B). Compared to the basal culture condition, the cUCB-MSCs had positive for Nestin, MAP2 with neuronal induction, but were negative prior to differentiation. These data showed that cUCB-MSCs had the ability to be inducted into glial and neuron cells under differentiation conditions (Figs. 2A-H).

Bottom Line: After neuronal differentiation, the cMSCs expressed the neuronal markers Nestin, GFAP, Tuj-1, microtubule-associated protein 2, NF160.With osteogenic differentiation, the cMSCs presented osteoblastic differentiation genes by RT-PCR.This finding also suggests that cMSCs might have the ability to differentiate multipotentially.

View Article: PubMed Central - PubMed

Affiliation: Adult Stem Cell Research Center, Department of Veterinary Public Health, College of Veterinery Medicine, Seoul National University, Seoul 151-742, Korea.

ABSTRACT
Human umbilical cord blood-derived mesenchymal stem cells (MSCs) are known to possess the potential for multiple differentiations abilities in vitro and in vivo. In canine system, studying stem cell therapy is important, but so far, stem cells from canine were not identified and characterized. In this study, we successfully isolated and characterized MSCs from the canine umbilical cord and its fetal blood. Canine MSCs (cMSCs) were grown in medium containing low glucose DMEM with 20% FBS. The cMSCs have stem cells expression patterns which are concerned with MSCs surface markers by fluorescence- activated cell sorter analysis. The cMSCs had multipotent abilities. In the neuronal differentiation study, the cMSCs expressed the neuronal markers glial fibrillary acidic protein (GFAP), neuronal class III beta tubulin (Tuj-1), neurofilament M (NF160) in the basal culture media. After neuronal differentiation, the cMSCs expressed the neuronal markers Nestin, GFAP, Tuj-1, microtubule-associated protein 2, NF160. In the osteogenic & chondrogenic differentiation studies, cMSCs were stained with alizarin red and toluidine blue staining, respectively. With osteogenic differentiation, the cMSCs presented osteoblastic differentiation genes by RT-PCR. This finding also suggests that cMSCs might have the ability to differentiate multipotentially. It was concluded that isolated MSCs from canine cord blood have multipotential differentiation abilities. Therefore, it is suggested that cMSCs may represent a be a good model system for stem cell biology and could be useful as a therapeutic modality for canine incurable or intractable diseases, including spinal cord injuries in future regenerative medicine studies.

Show MeSH
Related in: MedlinePlus