Construction of an allogenic chimeric mouse model for the study of the behaviors of donor stem cells in vivo / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is essential to establish an animal model for the elucidation of the biological behaviors of stem cells in vivo. We constructed a chimeric animal model by in utero transplantation for investigation of stem cell transplantation.</p><p><b>METHODS</b>This chimerism was achieved by injecting the stem cells derived from the bone marrow of green fluorescence protein (GFP)-transgenic mice into fetal mice at 13.5 days of gestation. Several methods such as polymerase chain reaction (PCR), real-time PCR, fluorescence-assisted cell sorting (FACS) and fluorescence in situ hybridization (FISH) were used for the observation of donor cells.</p><p><b>RESULTS</b>Under a fluorescence microscope, we observed the GFP cells of donor-origin in a recipient. PCR, FACS analysis and FISH indicated chimerism at various intervals. Real-time PCR indicated that some donor cells existed in chimera for more than 6 months.</p><p><b>CONCLUSIONS</b>Allogenic stem cells may exist in recipients for a long time and this allogenic animal model provides a useful tool for studying the behavior of hematopoietic stem cells and also offers an effective model system for the study of stem cells.</p>

The study on the stable inheritance and expression of foreign gene in transgenic mice / 生物工程学报

Two transgenic mouse strains, in which the expression of human factor IX (hFIX) in the milk were different significantly, were bred, and the foreign gene integration as well as the content of hFIX in the milk were detected by PCR, Southern blot, FISH and ELISA, respectively. The results showed that approximately 50% offsprings were transgenic positive. Foreign gene integrated in mouse chromosomes was intact. The hFIX expression of each mouse in the same strain was different, the content of hFIX in the milk was (43.32 +/- 5.41) microgram/mL in FIX-33 transgenic strain and (1.16 +/- 0.45) microgram/mL in FIX-124 transgenic strain. Meanwhile, the hFIX gene expression between the two strains was different remarkably (P < 0.01). We conclude that the characteristics of inheritance and expression in the founder were able to be transferred to their offsprings stably.