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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

Expression of GFP in human ADSCs.Human ADSCs were isolated from human adipose tissues and transduced with lentivirus vectors carrying GFP reporter gene. (A) spindle-like appearance of primary ADSCs at passage 3. (B) Phase-contrast microscopic image. (C) Fluorescence microscopic image. (D) merged image of (B) and (C). Original magnification of all images: 40 ×. (E) The effect of GFP expression on the proliferation of ADSCs.
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pone.0123264.g002: Expression of GFP in human ADSCs.Human ADSCs were isolated from human adipose tissues and transduced with lentivirus vectors carrying GFP reporter gene. (A) spindle-like appearance of primary ADSCs at passage 3. (B) Phase-contrast microscopic image. (C) Fluorescence microscopic image. (D) merged image of (B) and (C). Original magnification of all images: 40 ×. (E) The effect of GFP expression on the proliferation of ADSCs.

Mentions: Human primary ADSCs were obtained from human adipose tissues, and they demonstrated a fibroblast-like spindle shape (Fig 2A), similar to bone marrow- derived mesenchymal stem cells, and proliferated quickly in the culture medium. Before cell transplantation, ADSCs were transduced with lentivirus vector system carrying GFP reporter gene prior. The results indicated that more than 90% of ADSCs were positive for GFP expression after transduction and screening (Fig 2B–2D). In addition, exogenous GFP expression in ADSCs did not change cell morphology and cell viability (Fig 2E), which was in according with previous reports [29].


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)

Expression of GFP in human ADSCs.Human ADSCs were isolated from human adipose tissues and transduced with lentivirus vectors carrying GFP reporter gene. (A) spindle-like appearance of primary ADSCs at passage 3. (B) Phase-contrast microscopic image. (C) Fluorescence microscopic image. (D) merged image of (B) and (C). Original magnification of all images: 40 ×. (E) The effect of GFP expression on the proliferation of ADSCs.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0123264.g002: Expression of GFP in human ADSCs.Human ADSCs were isolated from human adipose tissues and transduced with lentivirus vectors carrying GFP reporter gene. (A) spindle-like appearance of primary ADSCs at passage 3. (B) Phase-contrast microscopic image. (C) Fluorescence microscopic image. (D) merged image of (B) and (C). Original magnification of all images: 40 ×. (E) The effect of GFP expression on the proliferation of ADSCs.
Mentions: Human primary ADSCs were obtained from human adipose tissues, and they demonstrated a fibroblast-like spindle shape (Fig 2A), similar to bone marrow- derived mesenchymal stem cells, and proliferated quickly in the culture medium. Before cell transplantation, ADSCs were transduced with lentivirus vector system carrying GFP reporter gene prior. The results indicated that more than 90% of ADSCs were positive for GFP expression after transduction and screening (Fig 2B–2D). In addition, exogenous GFP expression in ADSCs did not change cell morphology and cell viability (Fig 2E), which was in according with previous reports [29].

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