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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 structure of zebrafish embryo and experimental design.(A) Sketch of zebrafish embryo at selected stage. (B) The image of cells-injected dechorionated zebrafish embryo at 4.3 hpf. (C) We drew schematic representation of experimental design.
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pone.0123264.g001: The structure of zebrafish embryo and experimental design.(A) Sketch of zebrafish embryo at selected stage. (B) The image of cells-injected dechorionated zebrafish embryo at 4.3 hpf. (C) We drew schematic representation of experimental design.

Mentions: Wild-type zebrafish (Danio rerio) strain (AB) were raised and maintained according to standard procedures [28]. Each embryo chorion was carefully removed 3 h after fertilization with forceps under stereomicroscope. GFP-expressing human ADSCs were then transplanted into 200 zebrafish embryos at 3.3–4.3 hour post-fertilization (hpf) using glass needle under micromanipulator, as shown in Fig 1. Each embryo was injected with about 10 cells expressed GFP, and the control embryos did not receive cell transplantation. After transplantation, zebrafish embryos were maintained in 6-well plates for their development at 28°C, and observed using fluorescence microscope or laser confocal fluorescence microscope (Leica TCS SP8, Germany) at various time points.


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 structure of zebrafish embryo and experimental design.(A) Sketch of zebrafish embryo at selected stage. (B) The image of cells-injected dechorionated zebrafish embryo at 4.3 hpf. (C) We drew schematic representation of experimental design.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0123264.g001: The structure of zebrafish embryo and experimental design.(A) Sketch of zebrafish embryo at selected stage. (B) The image of cells-injected dechorionated zebrafish embryo at 4.3 hpf. (C) We drew schematic representation of experimental design.
Mentions: Wild-type zebrafish (Danio rerio) strain (AB) were raised and maintained according to standard procedures [28]. Each embryo chorion was carefully removed 3 h after fertilization with forceps under stereomicroscope. GFP-expressing human ADSCs were then transplanted into 200 zebrafish embryos at 3.3–4.3 hour post-fertilization (hpf) using glass needle under micromanipulator, as shown in Fig 1. Each embryo was injected with about 10 cells expressed GFP, and the control embryos did not receive cell transplantation. After transplantation, zebrafish embryos were maintained in 6-well plates for their development at 28°C, and observed using fluorescence microscope or laser confocal fluorescence microscope (Leica TCS SP8, Germany) at various time points.

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