Various types of somatic cells can be reprogrammed to induced pluripotent stem (iPS) cells. Somatic stem cells exhibit enhanced reprogramming efficiency by fewer factors, in contrast to fully differentiated cells. Nuclear LaminA is highly expressed in differentiated cells, and stem cells are characterized by the absence of LaminA. Granulosa cells (GCs) and cumulus cells in the ovarian follicles effectively and firstly generated cloned mice by somatic cell nuclear transfer, and these cells lack LaminA expression. We tested the hypothesis that GCs could be effectively used to generate iPS cells with fewer factors. We show that iPS cells are generated from GCs at high efficiency even with only two factors, Oct4 and Sox2, like the iPS cells generated using four Yamanaka factors. These iPS cells show pluripotency in vitro and in vivo, as evidenced by high expression of pluripotency-associated genes, Oct4, Nanog, and SSEA-1, differentiation into three embryonic germ layers by embryoid body formation and teratoma tests, as well as high efficient generation of chimeras. Moreover, the exogenous genes are effectively silenced in these iPS cells. These data provide additional evidence in supporting the notion that reduced expression of LaminA and stem cells can improve the reprogramming efficiency to pluripotency.
Get full access to this article
View all access options for this article.
References
1.
TakahashiK and YamanakaS. (2006). Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 126:663–676.
2.
MeyerN and PennLZ. (2008). Reflecting on 25 years with MYC. Nat Rev Cancer, 8:976–990.
3.
RowlandBD and PeeperDS. (2006). KLF4, p21 and context-dependent opposing forces in cancer. Nat Rev Cancer, 6:11–23.
4.
OkitaK, IchisakaT and YamanakaS. (2007). Generation of germline-competent induced pluripotent stem cells. Nature, 448:313–317.
5.
LiY, ZhangQ, YinX, YangW, DuY, et al. (2011). Generation of iPSCs from mouse fibroblasts with a single gene, Oct4, and small molecules. Cell Res, 21:196–204.
6.
ChenJ, LiuJ, YangJ, ChenY, NiS, et al. (2011). BMPs functionally replace Klf4 and support efficient reprogramming of mouse fibroblasts by Oct4 alone. Cell Res, 21:205–212.
7.
ZhuS, LiW, ZhouH, WeiW, AmbasudhanR, et al. (2010). Reprogramming of human primary somatic cells by OCT4 and chemical compounds. Cell Stem Cell, 7:651–655.
8.
ShiY, DoJT, DespontsC, HahmHS, ScholerHR, et al. (2008). A combined chemical and genetic approach for the generation of induced pluripotent stem cells. Cell Stem Cell, 2:525–528.
9.
TakahashiK, TanabeK, OhnukiM, NaritaM, IchisakaT, et al. (2007). Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 131:861–872.
10.
AoiT, YaeK, NakagawaM, IchisakaT, OkitaK, et al. (2008). Generation of pluripotent stem cells from adult mouse liver and stomach cells. Science, 321:699–702.
11.
LohYH, AgarwalS, ParkIH, UrbachA, HuoH, et al. (2009). Generation of induced pluripotent stem cells from human blood. Blood, 113:5476–5479.
12.
AasenT, RayaA, BarreroMJ, GarretaE, ConsiglioA, et al. (2008). Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes. Nat Biotechnol, 26:1276–1284.
13.
HaaseA, OlmerR, SchwankeK, WunderlichS, MerkertS, et al. (2009). Generation of induced pluripotent stem cells from human cord blood. Cell Stem Cell, 5:434–441.
14.
GiorgettiA, MontserratN, AasenT, GonzalezF, Rodriguez-PizaI, et al. (2009). Generation of induced pluripotent stem cells from human cord blood using OCT4 and SOX2. Cell Stem Cell, 5:353–357.
YanX, QinH, QuC, TuanRS, ShiS, et al. (2010). iPS cells reprogrammed from human mesenchymal-like stem/progenitor cells of dental tissue origin. Stem Cells Dev, 19:469–480.
17.
OdaY, YoshimuraY, OhnishiH, TadokoroM, KatsubeY, et al. (2010). Induction of pluripotent stem cells from human third molar mesenchymal stromal cells. J Biol Chem, 285:29270–29278.
18.
BrownME, RondonE, RajeshD, MackA, LewisR, et al. (2010). Derivation of induced pluripotent stem cells from human peripheral blood T lymphocytes. PLoS ONE, 5:e11373.
19.
LohYH, HartungO, LiH, GuoC, SahalieJM, et al. (2010). Reprogramming of T cells from human peripheral blood. Cell Stem Cell, 7:15–19.
20.
SekiT, YuasaS, OdaM, EgashiraT, YaeK, et al. (2010). Generation of induced pluripotent stem cells from human terminally differentiated circulating T cells. Cell Stem Cell, 7:11–14.
21.
StaerkJ, DawlatyMM, GaoQ, MaetzelD, HannaJ, et al. (2010). Reprogramming of human peripheral blood cells to induced pluripotent stem cells. Cell Stem Cell, 7:20–24.
22.
HannaJ, MarkoulakiS, SchorderetP, CareyBW, BeardC, et al. (2008). Direct reprogramming of terminally differentiated mature B lymphocytes to pluripotency. Cell, 133:250–264.
23.
PereiraCF, TerranovaR, RyanNK, SantosJ, MorrisKJ, et al. (2008). Heterokaryon-based reprogramming of human B lymphocytes for pluripotency requires Oct4 but not Sox2. PLoS Genet, 4:e1000170.
24.
EminliS, FoudiA, StadtfeldM, MaheraliN, AhfeldtT, et al. (2009). Differentiation stage determines potential of hematopoietic cells for reprogramming into induced pluripotent stem cells. Nat Genet, 41:968–976.
25.
EminliS, UtikalJ, ArnoldK, JaenischR and HochedlingerK. (2008). Reprogramming of neural progenitor cells into induced pluripotent stem cells in the absence of exogenous Sox2 expression. Stem Cells, 26:2467–2474.
26.
KimJB, ZaehresH, WuG, GentileL, KoK, et al. (2008). Pluripotent stem cells induced from adult neural stem cells by reprogramming with two factors. Nature, 454:646–650.
27.
GiorgettiA, MontserratN, Rodriguez-PizaI, AzquetaC, VeigaA, et al. (2010). Generation of induced pluripotent stem cells from human cord blood cells with only two factors: Oct4 and Sox2. Nat Protoc, 5:811–820.
28.
BlellochR, WangZ, MeissnerA, PollardS, SmithA, et al. (2006). Reprogramming efficiency following somatic cell nuclear transfer is influenced by the differentiation and methylation state of the donor nucleus. Stem Cells, 24:2007–2013.
29.
ZuoB, YangJ, WangF, WangL, YinY, et al. (2012). Influences of lamin A levels on induction of pluripotent stem cells. Biol Open, 1:1118–1127.
30.
ChronowskaE. (2012). Stem cell characteristics of ovarian granulosa cells—review. Ann Anim Sci, 12:151–157.
31.
GeL, SuiH-S, LanG-C, LiuN, WangJ-Z, et al. (2008). Coculture with cumulus cells improves maturation of mouse oocytes denuded of the cumulus oophorus: observations of nuclear and cytoplasmic events. Fertil Steril, 90:2376–2388.
32.
Kossowska-TomaszczukK, De GeyterC, De GeyterM, MartinI, HolzgreveW, et al. (2009). The multipotency of luteinizing granulosa cells collected from mature ovarian follicles. Stem Cells, 27:210–219.
33.
Kossowska-TomaszczukK and De GeyterC. (2013). Cells with stem cell characteristics in somatic compartments of the ovary. Biomed Res Int, 2013:310859.
34.
WakayamaT, PerryACF, ZuccottiM, JohnsonKR and YanagimachiR. (1998). Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature, 394:369–374.
35.
WellsDN, MisicaPM and TervitHR. (1999). Production of cloned calves following nuclear transfer with cultured adult mural granulosa cells. Biol Reprod, 60:996–1005.
36.
ParkK-W, KühholzerB, LaiL, MachátyZ, SunQ-Y, et al. (2001). Development and expression of the green fluorescent protein in porcine embryos derived from nuclear transfer of transgenic granulosa-derived cells. Anim Reprod Sci, 68:111–120.
37.
PolejaevaIA, ChenS-H, VaughtTD, PageRL, MullinsJ, et al. (2000). Cloned pigs produced by nuclear transfer from adult somatic cells. Nature, 407:86–90.
38.
GibbonsJ, AratS, RzucidloJ, MiyoshiK, WaltenburgR, et al. (2002). Enhanced survivability of cloned calves derived from roscovitine-treated adult somatic cells. Biol Reprod, 66:895–900.
39.
KeeferCL, KeystonR, LazarisA, BhatiaB, BeginI, et al. (2002). Production of cloned goats after nuclear transfer using adult somatic cells. Biol Reprod, 66:199–203.
40.
HuangJ, WangF, OkukaM, LiuN, JiG, et al. (2011). Association of telomere length with authentic pluripotency of ES/iPS cells. Cell Res, 21:779–792.
41.
WangF, YinY, YeX, LiuK, ZhuH, et al. (2012). Molecular insights into the heterogeneity of telomere reprogramming in induced pluripotent stem cells. Cell Res, 22:757–768.
42.
BarakatTS and GribnauJ. (2012). X chromosome inactivation in the cycle of life. Development, 139:2085–2089.
43.
MaheraliN, SridharanR, XieW, UtikalJ, EminliS, et al. (2007). Directly reprogrammed fibroblasts show global epigenetic remodeling and widespread tissue contribution. Cell Stem Cell, 1:55–70.
44.
MeissnerA, WernigM and JaenischR. (2007). Direct reprogramming of genetically unmodified fibroblasts into pluripotent stem cells. Nat Biotechnol, 25:1177–1181.
45.
StavropoulosN, LuN and LeeJT. (2001). A functional role for Tsix transcription in blocking Xist RNA accumulation but not in X-chromosome choice. Proc Natl Acad Sci U S A, 98:10232–10237.
46.
PoloJM, AnderssenE, WalshRM, SchwarzBA, NefzgerCM, et al. (2012). A molecular roadmap of reprogramming somatic cells into iPS cells. Cell, 151:1617–1632.
47.
SridharanR, TchieuJ, MasonMJ, YachechkoR, KuoyE, et al. (2009). Role of the murine reprogramming factors in the induction of pluripotency. Cell, 136:364–377.
48.
BuganimY, FaddahDA, ChengAW, ItskovichE, MarkoulakiS, et al. (2012). Single-cell expression analyses during cellular reprogramming reveal an early stochastic and a late hierarchic phase. Cell, 150:1209–1222.
49.
KimJB, SebastianoV, WuG, Araúzo-BravoMJ, SasseP, et al. (2009). Oct4-induced pluripotency in adult neural stem cells. Cell, 136:411–419.
50.
MontserratN, Ramirez-BajoMJ, XiaY, Sancho-MartinezI, Moya-RullD, et al. (2012). Generation of induced pluripotent stem cells from human renal proximal tubular cells with only two transcription factors, OCT4 and SOX2. J Biol Chem, 287:24131–24138.
51.
NakagawaM, TakizawaN, NaritaM, IchisakaT and YamanakaS. (2010). Promotion of direct reprogramming by transformation-deficient Myc. Proc Natl Acad Sci U S A, 107:14152–14157.
52.
NakagawaM, KoyanagiM, TanabeK, TakahashiK, IchisakaT, et al. (2008). Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat Biotechnol, 26:101–106.
53.
MiuraK, OkadaY, AoiT, OkadaA, TakahashiK, et al. (2009). Variation in the safety of induced pluripotent stem cell lines. Nat Biotechnol, 27:743–745.
54.
HuangfuD, MaehrR, GuoW, EijkelenboomA, SnitowM, et al. (2008). Induction of pluripotent stem cells by defined factors is greatly improved by small-molecule compounds. Nat Biotechnol, 26:795–797.
55.
MikkelsenTS, HannaJ, ZhangX, KuM, WernigM, et al. (2008). Dissecting direct reprogramming through integrative genomic analysis. Nature, 454:49–55.
56.
HuangfuD, OsafuneK, MaehrR, GuoW, EijkelenboomA, et al. (2008). Induction of pluripotent stem cells from primary human fibroblasts with only Oct4 and Sox2. Nat Biotechnol, 26:1269–1275.
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.
