Generation of C57BL/6 Knockout Mice Using C3H × BALB/c Blastocysts

Pacholczyk, Gabriela; Suhag, Rupali; Mazurek, Magdalena; Dederscheck, Suzanne M.; Koni, Pandelakis A.

doi:10.2144/000112706

Cited by 11 publications

(9 citation statements)

References 5 publications

(7 reference statements)

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“…Conventional microinjection into inbred or outbred albino blastocysts has been the standard method of injection with ES cells derived from the B6 strain (Lemckert et al 1997;Auerbach et al 2000;Schuster-Gossler et al 2001;Poueymirou et al 2007;Zevnik et al 2014), although the injection of 8-cell embryos increased the contribution of B6 ES cells to the resulting chimeras (Poueymirou et al 2007). Aggregation of B6 ES cells with outbred host morulae has been demonstrated to produce similar results without the need for sophisticated microinjection equipment (Auerbach et al 2000;Gertsenstein et al 2010), and various host embryo alternatives have been used in efforts to improve the efficiency of generating genetically modified mouse models (Pacholczyk et al 2008;Taft et al 2013;Zevnik et al 2014). In general, BALB/c and B6-albino inbred strains have been found to be more effective host embryos compared with other inbred or outbred strains for producing chimeras by conventional injection of B6 ES cells into blastocysts (Ledermann and Burki 1991;Auerbach et al 2000;Schuster-Gossler et al 2001).…”

Section: Introductionmentioning

confidence: 99%

Comparison of BALB/c and B6-albino mouse strain blastocysts as hosts for the injection of C57BL6/N-derived C2 embryonic stem cells

Alcantar¹,

Wiler²,

Rairdan³

2016

Transgenic Res

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Embryonic stem (ES) cells from a C57BL/6N (B6N) background injected into B6(Cg)-Tyrc-2J/J (B6-albino) recipient blastocysts are commonly used for generating genetically modified mouse models. To understand the influence of the recipient blastocyst strain on germline transmission, BALB/cAnNTac and B6-albino germline transmission rates were compared using the C57BL6/N-derived C2 ES cell line. A total of 92 ES cell clones from 27 constructs were injected. We compared blastocyst yield, birth rate, chimera formation rate, and high-percentage (>50 %) male chimera formation rate. For germline transmission, we analyzed 24 clones from 19 constructs, which generated high-percentage male chimeras from both donor strains. B6-albino hosts resulted in higher mean blastocyst yields per donor than did BALB/c ones (3.6 vs. 2.5). However, BALB/c hosts resulted in a higher birth rate than B6-albino ones (36 vs. 27 %), a higher chimera formation rate (50 vs. 42 %), a higher high-percentage male chimera rate (10 vs. 8 %), and a higher germline transmission rate (65 vs. 49 %), respectively. Our data suggest that BALB/c is a suitable blastocyst host strain for C2 ES cells and has an advantage over the B6-albino strain for receiving the injection of C2 ES cells.

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Section: Introductionmentioning

confidence: 99%

Comparison of BALB/c and B6-albino mouse strain blastocysts as hosts for the injection of C57BL6/N-derived C2 embryonic stem cells

Alcantar¹,

Wiler²,

Rairdan³

2016

Transgenic Res

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“…One of the best tools we have in understanding the biology of fibrosis is the use of genetically modified mice. Many genetically modified mice have been generated on a C57BL/6 background [1,2] due to several significant advantages of this strain [3]. However, ESKF research is hampered by a lack of both aggressive and relevant models of CKD in mice and especially in the fibrosis resistant C57BL/6 strain.…”

Section: Introductionmentioning

confidence: 99%

Development of a Chronic Kidney Disease Model in C57BL/6 Mice with Relevance to Human Pathology

Huang¹,

Scarpellini²,

Funck³

et al. 2013

Nephron Extra

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Background: Genetically modified mice are used to investigate disease and assess potential interventions. However, research into kidney fibrosis is hampered by a lack of models of chronic kidney disease (CKD) in mice. Recently, aristolochic acid nephropathy (AAN), characterised by severe tubulointerstitial fibrosis, has been identified as a cause of end stage kidney disease and proposed as a model of CKD. Published studies have used various dosing regimens, species and strains, with variable outcomes. Therefore, we aimed to develop a standardised protocol to develop tubulointerstitial fibrosis using pure aristolochic acid I (AAI) in C57BL/6 mice. Methods: AAI dose optimisation was performed by intraperitoneal injection of AAI at varying dose, frequency and duration. Kidney function was assessed by serum creatinine. Fibrosis was quantified by hydroxyproline levels and Masson’s Trichrome staining. Specific collagens were measured by immunofluorescent staining. Results: Single doses of AAI of >10 mg/kg caused acute kidney failure and death. Lower doses of 2.5 mg/kg needed to be administrated more than weekly to cause significant fibrosis. 3 mg/kg once every 3 days for 6 weeks followed by a disease development time of 6 weeks after AAI led to reduced kidney weight and function. Substantial tubulointerstitial fibrosis occurred, with males more severely affected. Increased deposition of collagen I, III and IV contributed to fibrosis, with collagen III and IV higher in males. Conclusions: AAN can be induced in C57BL/6 mice. The regimen of 3 mg/kg every 3 days for 6 weeks followed by 6 weeks of disease development time gives substantial tubulointerstitial fibrosis with lesions similar to those in humans.

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“…For these reasons, alternative host embryos for B6 ES cells have been investigated, e.g. C3H×BALB/c [28].…”

Section: Introductionmentioning

confidence: 99%

Efficient Generation of Germ Line Transmitting Chimeras from C57BL/6N ES Cells by Aggregation with Outbred Host Embryos

et al. 2010

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Genetically modified mouse strains derived from embryonic stem (ES) cells have become essential tools for functional genomics and biomedical research. Large scale mutagenesis projects are producing libraries of mutant C57BL/6 (B6) ES cells to enable the functional annotation of every gene of the mouse genome. To realize the utility of these resources, efficient and accessible methods of generating mutant mice from these ES cells are necessary. Here, we describe a combination of ICR morula aggregation and a chemically-defined culture medium with widely available and accessible components for the high efficiency generation of germline transmitting chimeras from C57BL/6N ES cells. Together these methods will ease the access of the broader biomedical research community to the publicly available B6 ES cell resources.

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Generation of C57BL/6 Knockout Mice Using C3H × BALB/c Blastocysts

Cited by 11 publications

References 5 publications

Comparison of BALB/c and B6-albino mouse strain blastocysts as hosts for the injection of C57BL6/N-derived C2 embryonic stem cells

Comparison of BALB/c and B6-albino mouse strain blastocysts as hosts for the injection of C57BL6/N-derived C2 embryonic stem cells

Development of a Chronic Kidney Disease Model in C57BL/6 Mice with Relevance to Human Pathology

Efficient Generation of Germ Line Transmitting Chimeras from C57BL/6N ES Cells by Aggregation with Outbred Host Embryos

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