Various factors affect the process of obtaining stable Arbas cashmere goat embryonic stem cells (ESCs), for example, the difficulty in isolating cells at the appropriate stage of embryonic development, the in vitro culture environment, and passage methods. With the emergence of induced pluripotent stem cell (iPSC) technology, it has become possible to use specific genes to induce somatic cell differentiation in PSCs. We transferred OCT4, SOX2, c-MYC, and KLF4 into Arbas cashmere goat fetal fibroblasts, then induced and cultured them using a drug-inducible system to obtain Arbas goat iPSCs that morphologically resembled mouse iPSCs. After identification, the obtained goat iPSCs expressed ESC markers, had a normal karyotype, could differentiate into embryoid bodies in vitro, and could differentiate into three germ layer cell types and form teratomas in vivo. We used microarray gene expression profile analysis to elucidate the reprogramming process. Our results provide the experimental basis for establishing cashmere goat iPSC lines and for future in-depth studies on molecular mechanism of cashmere goat somatic cell reprogramming.
Get full access to this article
View all access options for this article.
References
1.
BaoL., HeL., ChenJ., WuZ., LiaoJ., RaoL., RenJ., LiH., ZhuH., and QianL. (2011). Reprogramming of ovine adult fibroblasts to pluripotency via drug-inducible expression of defined factors. Cell Res., 21, 600–608.
2.
BehboodiE., BondarevaA., BeginI., RaoK., NeveuN., PiersonJ., WylieC., PieroF., HuangY., and ZengW. (2011). Establishment of goat embryonic stem cells from in vivo produced blastocyst‐stage embryos. Mol. Reprod. Dev., 78, 202–211.
3.
BrambrinkT., ForemanR., WelsteadG.G., LengnerC.J., WernigM., SuhH., and JaenischR. (2008). Sequential expression of pluripotency markers during direct reprogramming of mouse somatic cells. Cell Stem Cell, 2, 151–159.
4.
BuganimY., MarkoulakiS., van WietmarschenN., HokeH., WuT., GanzK., Akhtar-ZaidiB., HeY., AbrahamB.J., and PorubskyD. (2014). The developmental potential of iPSCs is greatly influenced by reprogramming factor selection. Cell Stem Cell, 15, 295–309.
5.
CapecchiM.R. (1989). Altering the genome by homologous recombination. Science, 244, 1288–1292.
6.
DamakS., JayN.P., BarrellG.K., and BullockD.W. (1996). Targeting gene expression to the wool follicle in transgenic sheep. Nat. Biotechnol., 14, 181–184.
7.
DattenaM., ChessaB., LacerenzaD., AccardoC., PilichiS., MaraL., ChessaF., VincentiL., and CappaiP. (2006). Isolation, culture, and characterization of embryonic cell lines from vitrified sheep blastocysts. Mol. Reprod. Dev., 73, 31–39.
8.
DoetschmanT., WilliamsP., and MaedaN. (1988). Establishment of hamster blastocyst-derived embryonic stem (ES) cells. Dev. Biol., 127, 224–227.
9.
DuY., WangJ., JiaJ., SongN., XiangC., XuJ., HouZ., SuX., LiuB., JiangT., ZhaoD., SunY., ShuJ., GuoQ., YinM., SunD., LuS., ShiY., and DengH. (2014). Human hepatocytes with drug metabolic function induced from fibroblasts by lineage reprogramming. Cell Stem Cell, 14, 394–403.
10.
EstebanM.A., XuJ., YangJ., PengM., QinD., LiW., JiangZ., ChenJ., DengK., and ZhongM. (2009). Generation of induced pluripotent stem cell lines from Tibetan miniature pig. J. Biol. Chem., 284, 17634–17640.
11.
EstebanM.A., WangT., QinB., YangJ., QinD., CaiJ., LiW., WengZ., ChenJ., and NiS. (2010). Vitamin C enhances the generation of mouse and human induced pluripotent stem cells. Cell Stem Cell, 6, 71–79.
12.
EvansM.J., and KaufmanM.H. (1981). Establishment in culture of pluripotential cells from mouse embryos. Nature, 292, 154–156.
13.
EzashiT., TeluguB.P.V., AlexenkoA.P., SachdevS., SinhaS., and RobertsR.M. (2009). Derivation of induced pluripotent stem cells from pig somatic cells. Proc. Natl. Acad. Sci. USA, 106, 10993–10998.
14.
HannaJ., WernigM., MarkoulakiS., SunC.-W., MeissnerA., CassadyJ.P., BeardC., BrambrinkT., WuL.-C., and TownesT.M. (2007). Treatment of sickle cell anemia mouse model with iPS cells generated from autologous skin. Science, 318, 1920–1923.
15.
HuangfuD., OsafuneK., MaehrR., GuoW., EijkelenboomA., ChenS., MuhlesteinW., and MeltonD.A. (2008). Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nat. Biotechnol., 26, 1269–1275.
16.
KimK., LerouP., YabuuchiA., LengerkeC., NgK., WestJ., KirbyA., DalyM.J., and DaleyG.Q. (2007). Histocompatible embryonic stem cells by parthenogenesis. Science, 315, 482–486.
17.
LiW., and DingS. (2010). Small molecules that modulate embryonic stem cell fate and somatic cell reprogramming. Trends Pharmacol. Sci., 31, 36–45.
18.
LiY., CangM., LeeA.S., ZhangK., and LiuD. (2011). Reprogramming of sheep fibroblasts into pluripotency under a drug-inducible expression of mouse-derived defined factors. PloS One, 6, e15947.
19.
LiaoJ., CuiC., ChenS., RenJ., ChenJ., GaoY., LiH., JiaN., ChengL., and XiaoH. (2009). Generation of induced pluripotent stem cell lines from adult rat cells. Cell Stem Cell, 4, 11–15.
20.
LiuH., ZhuF., YongJ., ZhangP., HouP., LiH., JiangW., CaiJ., LiuM., and CuiK. (2008). Generation of induced pluripotent stem cells from adult rhesus monkey fibroblasts. Cell Stem Cell, 3, 587–590.
21.
MartinG.R. (1981). Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc. Natl. Acad. Sci. USA, 78, 7634–7638.
22.
MunsieM.J., MichalskaA.E., O'BrienC.M., TrounsonA.O., PeraM.F., and MountfordP.S. (2000). Isolation of pluripotent embryonic stem cells from reprogrammed adult mouse somatic cell nuclei. Curr. Biol., 10, 989–992.
PawarS., MalakarD., DeA., and AksheyY. (2009). Stem cell-like outgrowths from in vitro fertilized goat blastocysts. Indian J. Exp. Biol., 47, 635.
25.
RenJ., PakY., HeL., QianL., GuY., LiH., RaoL., LiaoJ., CuiC., and XuX. (2011). Generation of hircine-induced pluripotent stem cells by somatic cell reprogramming. Cell Res., 21, 849.
26.
ShamblottM.J., AxelmanJ., WangS., BuggE.M., LittlefieldJ.W., DonovanP.J., BlumenthalP.D., HugginsG.R., and GearhartJ.D. (1998). Derivation of pluripotent stem cells from cultured human primordial germ cells. Proc. Natl. Acad. Sci. USA, 95, 13726–13731.
TakahashiK., and YamanakaS. (2006). Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 126, 663–676.
29.
TengH.F., KuoY.L., LooM.R., LiC.L., ChuT.W., SuoH., LiuH.S., LinK.H., and ChenS.L. (2010). Valproic acid enhances Oct4 promoter activity in myogenic cells. J. Cell. Biochem., 110, 995–1004.
30.
ThomasK.R., and CapecchiM.R. (1987). Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell, 51, 503–512.
WallR.J., RexroadC.E.Jr, PowellA., ShamayA., McKnightR., and HennighausenL. (1996). Synthesis and secretion of the mouse whey acidic protein in transgenic sheep. Transgenic Res., 5, 67–72.
33.
WangQ., XuX., LiJ., LiuJ., GuH., ZhangR., ChenJ., KuangY., FeiJ., and JiangC. (2011). Lithium, an anti-psychotic drug, greatly enhances the generation of induced pluripotent stem cells. Cell Res., 21, 1424–1435.
34.
WernigM., LengnerC.J., HannaJ., LodatoM.A., SteineE., ForemanR., StaerkJ., MarkoulakiS., and JaenischR. (2008). A drug-inducible transgenic system for direct reprogramming of multiple somatic cell types. Nat. Biotechnol., 26, 916–924.
35.
WuZ., ChenJ., RenJ., BaoL., LiaoJ., CuiC., RaoL., LiH., GuY., and DaiH. (2009). Generation of pig induced pluripotent stem cells with a drug-inducible system. J. Mol. Cell Biol., 1, 46–54.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
