由于莫納什大學的科學家們第一次從成年豹組織中產(chǎn)生胚胎干細胞樣細胞,瀕危滅絕雪豹的存活看到了希望,。
在從未有誘導多能干細胞(iPS)以前,胚胎干細胞的許多有用的特性產(chǎn)生自貓科成員,。此突破為將來克隆而冷凍保存遺傳物質(zhì)和其他輔助再生技術(shù)提供了可能性,。
發(fā)表在Theriogenology上的研究是Rajneesh Verma博士項目的一部分,由Paul Verma博士指導,,他們均來自莫納什醫(yī)學研究所(MIMR),。莫納什大學婦產(chǎn)科學系的副教授Peter Temple-Smith和昆士蘭大學的教授Michael Holland也共同研究此項目,。
研究人員用取自澳洲新南威爾斯的摩哥動物園(Mogo Zoo)成年雪豹的耳組織樣本產(chǎn)生誘導多能干細胞。
Verma博士說,,此突破很重要,,因為獲得生殖細胞,或配子相當困難,,甚至是從囚禁動物中獲取,。
"在瀕危物種組織的超低溫保存方面有很多意義,但是為了有用的保護而這樣做,,精子和卵子是必需的,。"
"干細胞的威力就是,它們在機體內(nèi)可分化成所有類型的細胞,。這就意味著,,它們有潛力成為配子。事實上,,小鼠誘導多功能干細胞已經(jīng)產(chǎn)生完整的后代,,因此可能性是巨大的", Verma博士說,。
貓科物種保存突破的益處與生物多樣性都很明顯,。
"通過產(chǎn)生這些干細胞,我已在從瀕危滅絕動物成年組織中創(chuàng)造生殖細胞方面邁出了第一步,。在將來,,我們的目標是利用誘導多功能干細胞的潛力,創(chuàng)造下一代,。這將有助于挽救瀕危滅絕的物種",, Verma先生說。
雪豹是一種大型貓科動物,,原產(chǎn)于亞洲中部山區(qū),。他們的高海撥棲息地和膽怯天性使我們很難做出準確的種群統(tǒng)計,但據(jù)估計,,自然界有3500至7000只雪豹,,數(shù)量還在下降。
Verma先生童年時期在印度就對這種巨大的貓科動物很著迷,。
"我確實是追隨著自己的熱情在運用自己干細胞的專業(yè)知識來拯救這些動物,。我在貓科動物其他成員身上運用同樣的技術(shù),其他的,,包括孟加拉虎,、捷豹和山貓。"(生物谷bioon.com)
doi:10.1016/j.theriogenology.2011.09.022
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Inducing pluripotency in somatic cells from the snow leopard (Panthera uncia), an endangered felid
R. Verma, M.K. Holland, P. Temple-Smith, P.J. Verma
Abstract Induced pluripotency is a new approach to produce embryonic stem-like cells from somatic cells that provides a unique means to understand both pluripotency and lineage assignment. To investigate whether this technology could be applied to endangered species, where the limited availability of gametes makes production and research on embryonic stem cells difficult, we attempted generation of induced pluripotent stem (iPS) cells from snow leopard (Panthera uncia) fibroblasts by retroviral transfection with Moloney-based retroviral vectors (pMXs) encoding four factors (OCT4, SOX2, KLF4 and cMYC). This resulted in the formation of small colonies of cells, which could not be maintained beyond four passages (P4). However, addition of NANOG, to the transfection cocktail produced stable iPS cell colonies, which formed as early as D3. Colonies of cells were selected at D5 and expanded in vitro. The resulting cell line was positive for alkaline phosphatase (AP), OCT4, NANOG, and Stage-Specific embryonic Antigen-4 (SSEA-4) at P14. RT-PCR also confirmed that endogenousOCT4 and NANOG were expressed by snow leopard iPS cells from P4. All five human transgenes were transcribed at P4, but OCT4, SOX2 and NANOG transgenes were silenced as early as P14; therefore, reprogramming of the endogenous pluripotent genes had occurred. When injected into immune-deficient mice, snow leopard iPS cells formed teratomas containing tissues representative of the three germ layers. In conclusion, this was apparently the first derivation of iPS cells from the endangered snow leopard and the first report on induced pluripotency in felid species. Addition of NANOG to the reprogramming cocktail was essential for derivation of iPS lines in this felid. The iPS cells provided a unique source of pluripotent cells with utility in conservation through cryopreservation of genetics, as a source of reprogrammed donor cells for nuclear transfer or for directed differentiation to gametes in the future.