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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

The proliferation of transplanted ADSCs detected by immunohistochemical staining.The GFP-expressing human ADSCs were transplanted into the zebrafish embryos at 3.3–4.3hpf. The proliferation of transplanted cells were evaluated by immunohistochemical staining with rabbit anti-human Ki-67 monoclonal antibody at 6 days post fertilization. (A) Representative image of immunohistochemical staining demonstrated that the proliferating cells were Ki-67 positive staining (brownish, indicated by green arrow), suggesting that the transplanted human ADSCs could proliferate in the zebrafish. (B) The control that did not receive cell transplantation, was negative staining of Ki-67. Scale bar: 100 μm.
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pone.0123264.g006: The proliferation of transplanted ADSCs detected by immunohistochemical staining.The GFP-expressing human ADSCs were transplanted into the zebrafish embryos at 3.3–4.3hpf. The proliferation of transplanted cells were evaluated by immunohistochemical staining with rabbit anti-human Ki-67 monoclonal antibody at 6 days post fertilization. (A) Representative image of immunohistochemical staining demonstrated that the proliferating cells were Ki-67 positive staining (brownish, indicated by green arrow), suggesting that the transplanted human ADSCs could proliferate in the zebrafish. (B) The control that did not receive cell transplantation, was negative staining of Ki-67. Scale bar: 100 μm.

Mentions: Immunohistochemical staining by using rabbit anti-human Ki-67 monoclonal antibody indicated that the proliferating human ADSCs in vivo were Ki-67 positive staining (brownish, indicated by green arrow) (Fig 6A) at 6 days post fertilization, but no Ki-67 positive cells were observed in fish cells and the control zebrafish (Fig 6B). Moreover, as shown in Fig 7, immunofluorescence staining of whole-mount zebrafish further confirmed that the Ki-67 positive cells were those transplanted human cells which demonstrated green and red fluorescence in the merged images. These data suggest that the transplanted human ADSCs could proliferate in zebrafish.


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)

The proliferation of transplanted ADSCs detected by immunohistochemical staining.The GFP-expressing human ADSCs were transplanted into the zebrafish embryos at 3.3–4.3hpf. The proliferation of transplanted cells were evaluated by immunohistochemical staining with rabbit anti-human Ki-67 monoclonal antibody at 6 days post fertilization. (A) Representative image of immunohistochemical staining demonstrated that the proliferating cells were Ki-67 positive staining (brownish, indicated by green arrow), suggesting that the transplanted human ADSCs could proliferate in the zebrafish. (B) The control that did not receive cell transplantation, was negative staining of Ki-67. Scale bar: 100 μm.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0123264.g006: The proliferation of transplanted ADSCs detected by immunohistochemical staining.The GFP-expressing human ADSCs were transplanted into the zebrafish embryos at 3.3–4.3hpf. The proliferation of transplanted cells were evaluated by immunohistochemical staining with rabbit anti-human Ki-67 monoclonal antibody at 6 days post fertilization. (A) Representative image of immunohistochemical staining demonstrated that the proliferating cells were Ki-67 positive staining (brownish, indicated by green arrow), suggesting that the transplanted human ADSCs could proliferate in the zebrafish. (B) The control that did not receive cell transplantation, was negative staining of Ki-67. Scale bar: 100 μm.
Mentions: Immunohistochemical staining by using rabbit anti-human Ki-67 monoclonal antibody indicated that the proliferating human ADSCs in vivo were Ki-67 positive staining (brownish, indicated by green arrow) (Fig 6A) at 6 days post fertilization, but no Ki-67 positive cells were observed in fish cells and the control zebrafish (Fig 6B). Moreover, as shown in Fig 7, immunofluorescence staining of whole-mount zebrafish further confirmed that the Ki-67 positive cells were those transplanted human cells which demonstrated green and red fluorescence in the merged images. These data suggest that the transplanted human ADSCs could proliferate in zebrafish.

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