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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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Related in: MedlinePlus

Identification of the cumulative population doubling level (CPDL) and culture of canine umbilical cord blood (cUCB)-mesenchymal stem cells (MSCs). Cells were cultured in DMEM (with 20% FBS). A: Two bars in a graph indicate the CPDL increase. Both bars show a consistently increasing growth rate during the passages. Each bar increase originates from the CPDL cumulative values, which were two separated sampled cells. B: Phase-contrast image of cUCB-MSCs, ×200.
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Figure 1: Identification of the cumulative population doubling level (CPDL) and culture of canine umbilical cord blood (cUCB)-mesenchymal stem cells (MSCs). Cells were cultured in DMEM (with 20% FBS). A: Two bars in a graph indicate the CPDL increase. Both bars show a consistently increasing growth rate during the passages. Each bar increase originates from the CPDL cumulative values, which were two separated sampled cells. B: Phase-contrast image of cUCB-MSCs, ×200.

Mentions: We isolated cUCB-MSCs from canine umbilical cord blood following to the cell isolation & culture method. The cUCB-MSCs (1 × 106) were collected and assessed in a T-25 cell culture flask. The passaged cells were collected every 2 days to count the cell number. The CPDL was measured and calculated and drawn as a graph. A consistently increasing rate of growth of the cumulative population was seen. Cells were cultured and maintained until passage 11. Small colonized populations were observed at the early stages of culture and dissociated for passaging. For each of the passages 1 to 11, cells were cryopreserved for further passaging and experiments (Fig. 1A). The morphology of cells was spindle-shape and typical fibroblast-like shape (Fig. 1B)


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)

Identification of the cumulative population doubling level (CPDL) and culture of canine umbilical cord blood (cUCB)-mesenchymal stem cells (MSCs). Cells were cultured in DMEM (with 20% FBS). A: Two bars in a graph indicate the CPDL increase. Both bars show a consistently increasing growth rate during the passages. Each bar increase originates from the CPDL cumulative values, which were two separated sampled cells. B: Phase-contrast image of cUCB-MSCs, ×200.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: Identification of the cumulative population doubling level (CPDL) and culture of canine umbilical cord blood (cUCB)-mesenchymal stem cells (MSCs). Cells were cultured in DMEM (with 20% FBS). A: Two bars in a graph indicate the CPDL increase. Both bars show a consistently increasing growth rate during the passages. Each bar increase originates from the CPDL cumulative values, which were two separated sampled cells. B: Phase-contrast image of cUCB-MSCs, ×200.
Mentions: We isolated cUCB-MSCs from canine umbilical cord blood following to the cell isolation & culture method. The cUCB-MSCs (1 × 106) were collected and assessed in a T-25 cell culture flask. The passaged cells were collected every 2 days to count the cell number. The CPDL was measured and calculated and drawn as a graph. A consistently increasing rate of growth of the cumulative population was seen. Cells were cultured and maintained until passage 11. Small colonized populations were observed at the early stages of culture and dissociated for passaging. For each of the passages 1 to 11, cells were cryopreserved for further passaging and experiments (Fig. 1A). The morphology of cells was spindle-shape and typical fibroblast-like shape (Fig. 1B)

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