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Xenotransplantation of human adipose-derived stem cells in zebrafish embryos.

Li J, Zeng G, Qi Y, Tang X, Zhang J, Wu Z, Liang J, Shi L, Liu H, Zhang P - PLoS ONE (2015)

Bottom Line: The results indicated that human ADSCs did not change their cell viability and the expression levels of cell surface antigens after GFP transduction.The expression of CD105 was observable in the xenotransplanted ADSCs, but CD31 expression was undetectable.Therefore, our results indicate that human ADSCs xenotransplanted in the zebrafish embryos not only can survive and proliferate at across-species circumstance, but also seem to maintain their undifferentiation status in a short term.

View Article: PubMed Central - PubMed

Affiliation: Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong Province, China.

ABSTRACT
Zebrafish is a widely used animal model with well-characterized background in developmental biology. The fate of human adipose-derived stem cells (ADSCs) after their xenotransplantation into the developing embryos of zebrafish is unknown. Therefore, human ADSCs were firstly isolated, and then transduced with lentiviral vector system carrying a green fluorescent protein (GFP) reporter gene, and followed by detection of their cell viability and the expression of cell surface antigens. These GFP-expressing human ADSCs were transplanted into the zebrafish embryos at 3.3-4.3 hour post-fertilization (hpf). Green fluorescent signal, the proliferation and differentiation of human ADSCs in recipient embryos were respectively examined using fluorescent microscopy and immunohistochemical staining. The results indicated that human ADSCs did not change their cell viability and the expression levels of cell surface antigens after GFP transduction. Microscopic examination demonstrated that green fluorescent signals of GFP expressed in the transplanted cells were observed in the embryos and larva fish at post-transplantation. The positive staining of Ki-67 revealed the survival and proliferation of human ADSCs in fish larvae after transplantation. The expression of CD105 was observable in the xenotransplanted ADSCs, but CD31 expression was undetectable. Therefore, our results indicate that human ADSCs xenotransplanted in the zebrafish embryos not only can survive and proliferate at across-species circumstance, but also seem to maintain their undifferentiation status in a short term. This xenograft model of zebrafish embryos may provide a promising and useful technical platform for the investigation of biology and physiology of stem cells in vivo.

No MeSH data available.


Related in: MedlinePlus

GFP-expressing ADSCs in zebrafish were observed by using laser confocal fluorescence microscope.Human ADSCs expressed GFP were xenotransplanted into zebrafish embryos at 3.3–4.3 hpf after their chorions were removed, and followed by observation under laser confocal fluorescence microscope at indicated time. Each embryo was injected with about 10 cells, and the control did not receive cell transplantation. (A) The control did not demonstrate any green fluorescence. (B) The representative image captured under a laser confocal fluorescence microscope at 15 dpf, displayed the GFP distributions (white arrows indicated) of transplanted cells in the zebrafish. Original magnification: 100 ×.
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pone.0123264.g005: GFP-expressing ADSCs in zebrafish were observed by using laser confocal fluorescence microscope.Human ADSCs expressed GFP were xenotransplanted into zebrafish embryos at 3.3–4.3 hpf after their chorions were removed, and followed by observation under laser confocal fluorescence microscope at indicated time. Each embryo was injected with about 10 cells, and the control did not receive cell transplantation. (A) The control did not demonstrate any green fluorescence. (B) The representative image captured under a laser confocal fluorescence microscope at 15 dpf, displayed the GFP distributions (white arrows indicated) of transplanted cells in the zebrafish. Original magnification: 100 ×.

Mentions: A total of 200 zebrafish embryos were transplanted with GFP-expressing human ADSCs, and 178 embryos at 24 hpf were alive after transplantation (survival rate 89%). Another 200 embryos did not receive any transplantation as the controls, but their chorions were removed like the transplanted groups, and 188 embryos were found to be alive at 24 hpf (survival rate 94%). There was no significant difference in survival rates between the groups. GFP-expressing human ADSCs in zebrafish embryos were observed at different time after transplantation. As shown in Fig 4, no fluorescence was observed in the control embryos or fish larva (Fig 4A1–4D1); on the other hand, green dots standing for human ADSCs in embryo (Fig 4A2) or larva (Fig 4B2) could be clearly observed with fluorescence microscopy at 12 or 24 phf after transplantation. With embryo development, green dots became irregular and disperse, but green fluorescence in vivo was still observable at 48 (Fig 4C2) and 96 phf (Fig 4D2). With time extension, we found that the green fluorescence in vivo could be sustained for more than 15 days (Fig 5). These results suggest that the transplanted human ADSCs can survive in zebrafish embryo and larva.


Xenotransplantation of human adipose-derived stem cells in zebrafish embryos.

Li J, Zeng G, Qi Y, Tang X, Zhang J, Wu Z, Liang J, Shi L, Liu H, Zhang P - PLoS ONE (2015)

GFP-expressing ADSCs in zebrafish were observed by using laser confocal fluorescence microscope.Human ADSCs expressed GFP were xenotransplanted into zebrafish embryos at 3.3–4.3 hpf after their chorions were removed, and followed by observation under laser confocal fluorescence microscope at indicated time. Each embryo was injected with about 10 cells, and the control did not receive cell transplantation. (A) The control did not demonstrate any green fluorescence. (B) The representative image captured under a laser confocal fluorescence microscope at 15 dpf, displayed the GFP distributions (white arrows indicated) of transplanted cells in the zebrafish. Original magnification: 100 ×.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4388839&req=5

pone.0123264.g005: GFP-expressing ADSCs in zebrafish were observed by using laser confocal fluorescence microscope.Human ADSCs expressed GFP were xenotransplanted into zebrafish embryos at 3.3–4.3 hpf after their chorions were removed, and followed by observation under laser confocal fluorescence microscope at indicated time. Each embryo was injected with about 10 cells, and the control did not receive cell transplantation. (A) The control did not demonstrate any green fluorescence. (B) The representative image captured under a laser confocal fluorescence microscope at 15 dpf, displayed the GFP distributions (white arrows indicated) of transplanted cells in the zebrafish. Original magnification: 100 ×.
Mentions: A total of 200 zebrafish embryos were transplanted with GFP-expressing human ADSCs, and 178 embryos at 24 hpf were alive after transplantation (survival rate 89%). Another 200 embryos did not receive any transplantation as the controls, but their chorions were removed like the transplanted groups, and 188 embryos were found to be alive at 24 hpf (survival rate 94%). There was no significant difference in survival rates between the groups. GFP-expressing human ADSCs in zebrafish embryos were observed at different time after transplantation. As shown in Fig 4, no fluorescence was observed in the control embryos or fish larva (Fig 4A1–4D1); on the other hand, green dots standing for human ADSCs in embryo (Fig 4A2) or larva (Fig 4B2) could be clearly observed with fluorescence microscopy at 12 or 24 phf after transplantation. With embryo development, green dots became irregular and disperse, but green fluorescence in vivo was still observable at 48 (Fig 4C2) and 96 phf (Fig 4D2). With time extension, we found that the green fluorescence in vivo could be sustained for more than 15 days (Fig 5). These results suggest that the transplanted human ADSCs can survive in zebrafish embryo and larva.

Bottom Line: The results indicated that human ADSCs did not change their cell viability and the expression levels of cell surface antigens after GFP transduction.The expression of CD105 was observable in the xenotransplanted ADSCs, but CD31 expression was undetectable.Therefore, our results indicate that human ADSCs xenotransplanted in the zebrafish embryos not only can survive and proliferate at across-species circumstance, but also seem to maintain their undifferentiation status in a short term.

View Article: PubMed Central - PubMed

Affiliation: Institute of Plastic Surgery, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong Province, China.

ABSTRACT
Zebrafish is a widely used animal model with well-characterized background in developmental biology. The fate of human adipose-derived stem cells (ADSCs) after their xenotransplantation into the developing embryos of zebrafish is unknown. Therefore, human ADSCs were firstly isolated, and then transduced with lentiviral vector system carrying a green fluorescent protein (GFP) reporter gene, and followed by detection of their cell viability and the expression of cell surface antigens. These GFP-expressing human ADSCs were transplanted into the zebrafish embryos at 3.3-4.3 hour post-fertilization (hpf). Green fluorescent signal, the proliferation and differentiation of human ADSCs in recipient embryos were respectively examined using fluorescent microscopy and immunohistochemical staining. The results indicated that human ADSCs did not change their cell viability and the expression levels of cell surface antigens after GFP transduction. Microscopic examination demonstrated that green fluorescent signals of GFP expressed in the transplanted cells were observed in the embryos and larva fish at post-transplantation. The positive staining of Ki-67 revealed the survival and proliferation of human ADSCs in fish larvae after transplantation. The expression of CD105 was observable in the xenotransplanted ADSCs, but CD31 expression was undetectable. Therefore, our results indicate that human ADSCs xenotransplanted in the zebrafish embryos not only can survive and proliferate at across-species circumstance, but also seem to maintain their undifferentiation status in a short term. This xenograft model of zebrafish embryos may provide a promising and useful technical platform for the investigation of biology and physiology of stem cells in vivo.

No MeSH data available.


Related in: MedlinePlus