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KMID : 0366220160510010031
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Korean Journal of Hematology
2016 Volume.51 No. 1 p.31 ~ p.36
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Humanizing NOD/SCID/IL-2Rγnull (NSG) mice using busulfan and retro-orbital injection of umbilical cord blood-derived CD34+ cells
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Kang Young-Kyung
Ko Yun-Mi
Choi Ae-Ry
Lee Jun-Ah
Choi Hyeong-Jwa
Seo Jin-Hee
Lee Min-Young
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Abstract
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Background: Humanized mouse models are still under development, and various protocols exist to improve human cell engraftment and function.
Methods: Fourteen NOD/SCID/IL-2Rγnull (NSG) mice (4?5 wk old) were conditioned with busulfan and injected with human umbilical cord blood (hUCB)-derived CD34+ hematopoietic stem cells (HSC) via retro-orbital sinuses. The bone marrow (BM), spleen, and peripheral blood (PB) were analyzed 8 and 12 weeks after HSC transplantation.
Results: Most of the NSG mice tolerated the regimen well. The percentage of hCD45+ and CD19+cells rose significantly in a time-dependent manner. The median percentage of hCD45+cells in the BM was 55.5% at week 8, and 67.2% at week 12. The median percentage of hCD45+ cells in the spleen at weeks 8 and 12 was 42% and 51%, respectively. The median percentage of hCD19+ cells in BM at weeks 8 and 12 was 21.5% and 39%, respectively (P=0.04). Similarly, the median percentage of hCD19+ cells in the spleen at weeks 8 and 12 was 10% and 24%, respectively (P=0.04). The percentage of hCD19+B cells in PB was 23% at week 12. At week 8, hCD3+ T cells were barely detectable, while hCD7+ was detected in the BM and spleen. The percentage of hCD3+ T cells was 2?3% at week 12 in the BM, spleen, and PB of humanized NSG mice.
Conclusion: We adopted a simplified protocol for establishing humanized NSG mice. We observed a higher engraftment rate of human CD45+ cells than earlier studies without any significant toxicity. And human CD45+ cell engraftment at week 8 was comparable to that of week 12.
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KEYWORD
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Humanized mice, Busulfan, Retro-orbital sinus, Hematopoietic stem cell
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