Bioluminescence imaging as a tool to evaluate germ cells in vitro and transplantation in vivo as fertility preservation of prepubertal male mice

Chen, Chi Huang; Wang, Chia-Woei; Hsu, Meng‐Ting; Huang, Yen Hua; Lai, Wen-FuThomas; Tzeng, Chii Ruey

doi:10.1016/j.fertnstert.2012.02.003

Cited by 9 publications

(15 citation statements)

References 22 publications

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“…For these experiments, they utilized SSCs isolated from Acr-GFP Tg mice as a reporter, which allowed them to detect the appearance of post-meiotic spermatids in real time. Meanwhile, a bioluminescence imaging, which has been widely utilized for cancer biology, is also reported to be feasible in germ cell study when it's combined with transplantation (Chen et al, 2012 ).…”

Section: Introductionmentioning

confidence: 99%

Generation of a dual-color reporter mouse line to monitor spermatogenesis in vivo

Makino

Inoue

Hada

et al. 2014

Front. Cell Dev. Biol.

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In vivo fluorescent imaging technique is a strong tool to visualize the various cellular events such as the proliferation, differentiation, migration, and a lineage tracing in living cells requiring no further experimental procedure such as immunostaining. During spermatogenesis, unique and dynamic histone exchanges occur. Since the timing and types of histone exchanges defines the particular stages of spermatogenesis, visualizing certain types of histones in testes is useful not only for researching specific histone dynamics, but also for monitoring the stages of spermatogenesis in vivo. In this study, we report the establishment of two transgenic (Tg) mouse lines expressing histone H4-Venus (H4V) and histone H3.3-mCherry (H33C) fusion proteins in testicular germ cells, and demonstrated their utility for monitoring germ cell development in vivo. Because of the choice of promoter as well as the nature of these histones, H4V and H33C were exclusively expressed in the germ cells of the distinct stages, which allowed the determination of spermatogenic stages in real time. In addition, disappearance of H4V and H33C at particular stages of differentiation/fertilization also represented dynamic histone removal. Collectively, these Tg mice are a valuable resource not only for monitoring differentiation stages, but also for studying the chromatin dynamics of post-natal testicular germ cell development in vivo.

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Section: Introductionmentioning

confidence: 99%

Generation of a dual-color reporter mouse line to monitor spermatogenesis in vivo

Makino

Inoue

Hada

et al. 2014

Front. Cell Dev. Biol.

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“…In our previous studies, we applied BLI as a tool to evaluate the effects of immunosuppression after allotransplantation of ovarian grafts [28], to track the rejection and survival of mouse ovarian iso- and allografts, and to evaluate germ cells in vitro and transplantation in vivo as a part of fertility preservation in prepubertal male mice [29,30]. …”

Section: Resultsmentioning

confidence: 99%

In vivo fate mapping of cryopreserved murine ovarian grafts

Chen

Tzeng

2014

J Ovarian Res

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BackgroundCryopreservation of ovarian tissue has been suggested as an alternative to restore fertility for ovarian failure before chemotherapy.MethodsOvaries of donor FVB/N-Tg (PolII–Luc) Ltc transgenic mice (n = 5) were cryopreserved and transplanted to the back muscles of recipient FVB/NJNarl wild-type mice that had undergone bilateral oophorectomy. We evaluated the fate of cryopreserved murine ovarian grafts by in vivo bioluminescent imaging (BLI), AMH mRNA expression and follicle counts.ResultsThere were significantly stronger BLI signals in the fresh ovaries than in the frozen–thawed ones. The number of primordial follicles was significantly lower in frozen–thawed ovaries at 10 days after transplantation (P < 0.001). The AMH mRNA expression was significantly lower in the frozen–thawed ovaries (P < 0.001), showing that unavoidable harm occurs after transplantation.ConclusionsOvarian cryopreservation by slow freezing compromises ovarian reserve by cryoinjury and ischemia, evident at an early stage after transplantation.

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“…The brightness and tissue penetration of the bioluminescent signal has limited its wide application in vivo. In previous studies 12,[21][22][23] , nearly all in vivo cell-tracking experiments with bioluminescence used a small animal (mouse or zebrafish, etc.) as research model.…”

Section: Discussionmentioning

confidence: 99%

Non-Invasive Cell Tracking with Brighter and Red-Transferred Luciferase for Potential Application in Stem Cell Therapy

Dou

Matz

et al. 2019

Cell Transplant

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This study investigated the safety of a novel cell-labeling technology with mKATE and Renilla reniformis luciferase (mKATE-renLUC) and assessed the efficacy on tracking implanted human placental stromal cells (PSC) in an erectile dysfunction (ED) animal model. Human PSC were labeled with mKATE-renLUC using a lentivirus. Cell viability, apoptosis, proliferation, migration, surface marker expression and differentiation potential of the labeled PSC were evaluated and compared with non-labeled PSC. The paracrine profile of labeled cells was examined using an angiogenesis protein array. The brightness and duration of labeled cells with different densities were evaluated. An ED rat model was established and labeled PSC were injected into cavernosal tissue of the penis. The migration and distribution of transplanted PSC were monitored using an IVIS imaging system in real time. Implanted PSC were identified in isolated tissues via detection of mKATE fluorescence. The cell viability, morphology, proliferation, migration, surface marker expression and differentiation potential of mKATE-renLUC-labeled PSC were similar to those of non-labeled cells in vitro (no statistical difference p>0.05). Similar expressions of trophic factors were found between labeled and non-labeled PSC. The migration and distribution of PSC expressing renLUC were tracked in vivo using IVIS imaging system. mKATE-positive PSC were detected in penile, kidney, prostate and hepatic tissues using histological methods. This labeling technology provides a safe and effective cell-tracking approach with a brighter fluorophore and codon-optimized luciferase.

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Bioluminescence imaging as a tool to evaluate germ cells in vitro and transplantation in vivo as fertility preservation of prepubertal male mice

Cited by 9 publications

References 22 publications

Generation of a dual-color reporter mouse line to monitor spermatogenesis in vivo

Generation of a dual-color reporter mouse line to monitor spermatogenesis in vivo

In vivo fate mapping of cryopreserved murine ovarian grafts

Non-Invasive Cell Tracking with Brighter and Red-Transferred Luciferase for Potential Application in Stem Cell Therapy

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