Generation of American mink induced pluripotent stem cells: a protocol

Пристяжнюк, И. Е.; Мензоров, А. Г.

doi:10.18699/vj17.288

Cited by 2 publications

(2 citation statements)

References 9 publications

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“…We estimated the transduction efficiency of the cell lines using a lentiviral vector with GFP according to Menzorov et al [22] and Pristyazhnyuk and Menzorov [23] with minor modifications. The lentiviruses with GFP were produced in the Phoenix cell line using Lipofectamine 3000 reagent (cat#L3000-008, Thermo Fisher Scientific).…”

Section: Transduction With Lentivirus Vectors Expressing Gfpmentioning

confidence: 99%

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Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies

Pristyazhnyuk

Malinovskaya

Бородин

2022

Animals

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The last decade was marked by a steep rise in avian studies at genomic and cellular levels. Cell lines are important tools for in vitro studies in cell biology and cytogenetics. We developed a simple method of primary somatic cell culture establishment from the ovaries of the great tits (Parus major) and testes of ten Passerine species, characterized the cellular composition of the ovary-derived lines using RT-PCR and immunolocalization of the tissue-specific markers and tested the efficiency of two methods of genetic transformation of the ovary-derived cell line. We found that the ovary-derived cell cultures of the great tit were composed of fibroblasts mainly, but also contained interstitial and granulosa cells. They were cultivated until the 10th passage without any noticeable decrease in their proliferative activity. The testis-derived cell cultures had lower proliferative potential. However, both ovary- and testis-derived cell cultures provided enough material for high quality mitotic metaphase chromosome preparations. The efficiency of its transduction with lentivirus containing a GFP reporter was very low, while electroporation with episomal vectors expressing GFP resulted in a high yield of GFP-positive cells. The proposed method could be used for the generation of high quality material for various cytogenetic and genomic studies.

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Section: Transduction With Lentivirus Vectors Expressing Gfpmentioning

confidence: 99%

“…The transduction was carried out with 10 or 50% lentivirus-containing supernatant in a growth medium without antibiotics with heat-inactivated FBS, chicken serum, and 10 µg/mL Polybrene (cat#TR-1003-G, Sigma-Aldrich). The multiplicity of infection (MOI) was estimated as previously described [23].…”

Section: Transduction With Lentivirus Vectors Expressing Gfpmentioning

confidence: 99%

Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies

Pristyazhnyuk

Malinovskaya

Бородин

2022

Animals

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Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts

Beklemisheva

Мензоров

2019

Vestn. VOGiS

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Generation of induced pluripotent stem (iPS) cells expanded possibilities of pluripotency and early development studies. Generation of order Carnivora iPS cells from dog (Canis lupus familiaris), snow leopard (Panthera uncia), and American mink (Neovison vison) was previously reported. The aim of the current study was to examine conditions of pinniped fbroblast reprogramming. Pinnipeds are representatives of the suborder Caniformia sharing conservative genomes. There are several ways to deliver reprogramming transcription factors: RNA, proteins, plasmids, viral vectors etc. The most eﬀective delivery systems for mouse and human cells are based on viral vectors. We compared a lentiviral vector which integrates into the genome and a Sendai virusbased vector, CytoTune EmGFP Sendai Fluorescence Reporter. The main advantage of Sendai virusbased vectors is that they do not integrate into the genome. We performed delivery of genetic constructions carrying ﬂuorescent proteins to fbroblasts of seven Pinnipeds: northern fur seal (Callorhinus ursinus), Steller sea lion (Eumetopias jubatus), walrus (Odobenus rosmarus), bearded seal (Erignathus barbatus), Baikal seal (Pusa sibirica), ringed seal (Phoca hispida), and spotted seal (Phoca largha). We also transduced American mink (N. vison), human (Homo sapiens), and mouse (Mus musculus) fbroblasts as a control. We showed that the Sendai virusbased transduction system provides transgene expression onetwo orders of magnitude higher than the lentiviral system at a comparable multiplicity of infection. Also, transgene expression after Sendai virusbased transduction is quite stable and changes only slightly at day four compared to day two. These data allow us to suggest that Sendai virusbased vectors are preferable for generation of Pinniped iPS cells.

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Generation of American mink induced pluripotent stem cells: a protocol

Cited by 2 publications

References 9 publications

Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies

Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies

Use of a Sendai virus-based vector for effcient transduction of pinniped fbroblasts

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