Development of a positive method for male stem cell-mediated gene transfer in mouse and pig

Kim, Jin‐Hoi; Jung-Ha, Haesook; Lee, Hoon-Taek; Chung, Kil-Saeng

doi:10.1002/(sici)1098-2795(199704)46:4<515::aid-mrd10>3.0.co;2-v

Cited by 79 publications

(47 citation statements)

References 40 publications

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“…In this way, transgenic spermatozoa-producing animals have been generated, but the yield (7-13%) was not comparable with the classical ex vivo SMGT technique (5-60%; Kim et al 1997). Very recently, the impact of electroporation treatment on the organism has also been assessed, showing that damage inflicted to spermatogenesis by electroporation can severely impair the production of sperm cells (Umemoto et al 2005).…”

Section: Modifications Of Sperm-mediated Gene Transfermentioning

confidence: 99%

Sperm-mediated gene transfer

Lavitrano

Busnelli

Cerrito

et al. 2006

Reprod. Fertil. Dev.

115

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Abstract. Since 1989, a new method for the production of transgenic animals has been available, namely spermmediated gene transfer (SMGT), based on the intrinsic ability of sperm cells to bind and internalise exogenous DNA molecules and to transfer them into the oocyte at fertilisation. We first described the SMGT procedure in a small animal model, with high efficiency reported in the mouse. In addition, we successfully adapted and optimised the technique for use in large animals; it was, in fact, highly efficient in the generation of human decay accelerating factor transgenic pig lines, as well as multigene transgenic pigs in which three different reporter genes, namely enhanced green fluorescent protein, enhanced blue fluorescent protein and red fluorescent protein, were introduced. The major benefits of the SMGT technique were found to be its high efficiency, low cost and ease of use compared with other methods. Furthermore, SMGT does not require embryo handling or expensive equipment. Sperm-mediated gene transfer could also be used to generate multigene transgenic pigs that would be of benefit as large animal models for medical research, for agricultural and pharmaceutical applications and, in particular, for xenotransplantation, which requires extensive genetic manipulation of donor pigs to make them suitable for grafting to humans.

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Section: Modifications Of Sperm-mediated Gene Transfermentioning

confidence: 99%

Sperm-mediated gene transfer

Lavitrano

Busnelli

Cerrito

et al. 2006

Reprod. Fertil. Dev.

115

View full text Add to dashboard Cite

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“…The required sterilizing dose reported in the literature varies widely between different species. For example, doses of 40-100 mg per kg body weight were used in pigs [19] and 4-12 mg kg −1 doses were used in coyotes [20]. Recipient rats were usually prepared by administering two injections of 10 or 15 mg kg −1 of busulfan [21][22], whereas 40-44 mg kg −1 of busulfan was the most commonly used dose for mice [1,3,12,23].…”

Section: Introductionmentioning

confidence: 99%

Optimal dose of busulfan for depleting testicular germ cells of recipient mice before spermatogonial transplantation

Wang¹,

Zhou²,

Yuan³

et al. 2009

Asian J Androl

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Successful spermatogonial transplantation requires depletion of the host germ cells to allow efficient colonization of the donor spermatogonial stem cells. Although a sterilizing drug, busulfan (Myleran), is commonly used for preparing a recipient mouse before transplantation, the optimal dose of this drug has not yet been defined. The present study investigated the effects of different doses of busulfan (10-50 mg per kg body weight) on survival rate, testicular mass and histomorphology, and on the haploid spermatids and spermatozoa of male BALB/c mice. The results suggest that a dosage of 30 mg kg -1 is optimal for the ablative treatment with busulfan used to prepare the recipient mice. This dose results in an adequate depletion of the host germ cells for colonization of donorderived spermatogonial stem cells and causes the lowest death rate of the animals.

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“…Spermatogonial stem cell-based methods of transferring genes to new generations provide a promising future in the field of animal science to accelerate the generation of transgenic animals more rapidly and effectively (Wang et al, 2014). However, to gain these desirable outcomes, some difficulties need to be surmounted (Kim et al, 1997).…”

Section: Introductionmentioning

confidence: 53%

Optimization of <i>in vitro</i> culture and transfection condition of bovine primary spermatogonial stem cells

Jafarnejad¹,

Aminafshar²,

Zandi³

et al. 2018

SA J. An. Sci.

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The present study aimed to optimize the in vitro culture and transfection efficiency of bovine primary spermatogonial stem cells (SSCs). To this end, SSCs were obtained from newborn Holstein bull calves by two-step enzymatic digestion. After enrichment and culture, SSCs were characterized by using alkaline phosphatase (AP) staining and expression of vasa and thy1 genes as specific bovine SSC markers. To evaluate the effect of antioxidants on vitality, colony formation, and the expression of pro-and anti-apoptotic genes of bovine SSCs, various concentrations of vitamin C (5, 10, 25 and 50 µg/mL) and Trolox (a water soluble α-tocopherol analogue) (12.5, 25, 50 and 100 µg/mL) were added to the SSC culture medium. The results showed that SSCs treated with 50 µg/mL of vitamin C or 25 µg/mL of Trolox individually could increase cell viability and colony formation significantly in comparison with other concentrations and the control group. Additionally, the expressions of bax (as a pro-apoptotic gene) and bcl2 (as an anti-apoptotic gene) were significantly lower and higher than the control group, respectively. To optimize the transfection condition, the effective dosages of vitamin C or Trolox, with various concentrations of two transfection reagents (X-tremeGENE HP and Turbofect) and DNA, at day 8 of culture, were studied. Results showed that 1 μl X-tremeGENE HP or 0.5 μl Turbofect and 2 μg of DNA are the best concentrations for transfecting SSCs. However, X-tremeGENE HP expressed more potential for transfecting SSCs in comparison with Turbofect. Besides, no difference was observed between the use of defined doses of vitamin C or Trolox. ____________________________________________________________________________________

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Development of a positive method for male stem cell-mediated gene transfer in mouse and pig

Cited by 79 publications

References 40 publications

Sperm-mediated gene transfer

Sperm-mediated gene transfer

Optimal dose of busulfan for depleting testicular germ cells of recipient mice before spermatogonial transplantation

Optimization of <i>in vitro</i> culture and transfection condition of bovine primary spermatogonial stem cells

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