Restricted accessResearch articleFirst published online 2021-06
Two Small Molecule Inhibitors Promote Reprogramming of Guangxi Bama Mini-Pig Mesenchymal Stem Cells Into Naive-Like State Induced Pluripotent Stem Cells
Past researches have shown that pluripotency maintenance of naive and primed-state pluripotent stem cells (PSCs) depends on different signaling pathways, and naive-state PSCs possess the ability to produce chimeras when they are introduced into a blastocyst. Considering porcine is an attractive model for preclinical studies, many researches about pig induced pluripotent stem cells (piPSCs) have been reported. Some cytokines and small molecule compounds could transform primed piPSCs into naive state. However, there are no suitable culture conditions for generation of naive-state piPSCs with high efficiency; other small molecule compounds need further exploration. In this study, we investigated whether p38 MAPK and JNK signal pathway inhibitor SB203580 and SP600125 could be of benefit for acquiring naive-state piPSCs. By comparing reprogramming efficiencies under conditions of different donor cells and culture environment, we found that porcine bone marrow mesenchymal stem cells (PBMSCs) have higher efficiency on piPSC induction, and the culture condition of CHIR99021+PD0325901(2i)+Lif+bFGF is more suitable for subculturing of piPSCs. Our results also indicate that SB203580 and SP600125 could promote reprogramming of PBMSCs into naive-like state piPSCs. These results provide guidance for choosing donor cells, culture conditions, and research of different state iPSCs during the process of reprogramming pig somatic cells.
Get full access to this article
View all access options for this article.
References
1.
AhmadzadehN., RoberingJ.W., Kengelbach-WeigandA., Al-AbboodiM., BeierJ.P., HorchR.E., and BoosA.M. (2020). Human adipose-derived stem cells support lymphangiogenesis in vitro by secretion of lymphangiogenic factors. Exp. Cell. Res. 388,111816.
2.
AltshulerA., VerbukM., BhattacharyaS., AbramovichI., HaklaiR., HannaJ.H., KloogY., GottliebE., and Shalom-FeuersteinR. (2018). RAS regulates the transition from naive to primed pluripotent stem cells. Stem Cell Rep. 10, 1088–1101.
3.
BourillotP.Y., SantamariaC., DavidL., and SavatierP. (2020). GP130 signaling and the control of naive pluripotency in humans, monkeys, and pigs. Exp. Cell. Res. 386,111712.
4.
ChaH.J., YunJ.I., HanN.R., KimH.Y., BaekS., LeeS.H., LeeJ., and LeeE. (2018). Generation of embryonic stem-like cells from in vivo-derived porcine blastocysts at a low concentration of basic fibroblast growth factor. Reprod. Domest. Anim. 53, 176–185.
5.
ChoiK.H., ParkJ.K., SonD., HwangJ.Y., LeeD.K., KaH., ParkJ., and LeeC.K. (2016). Reactivation of endogenous genes and epigenetic remodeling are barriers for generating transgene-free induced pluripotent stem cells in pig. PLoS One 11,e0158046.
6.
DengY., LiuQ., LuoC., ChenS., LiX., WangC., LiuZ., LeiX., ZhangX., SunH., et al. (2012). Generation of induced pluripotent stem cells from buffalo (Bubalus bubalis) fetal fibroblasts with buffalo defined factors. Stem Cells Dev. 21, 2485–2494.
7.
DengY., HuangG., ChenF., TestroetE.D., LiH., LiH., NongT., YangX., CuiJ., ShiD., and YangS. (2019). Hypoxia enhances buffalo adipose-derived mesenchymal stem cells proliferation, stemness, and reprogramming into induced pluripotent stem cells. J. Cell. Physiol. 234, 17254–17268.
8.
DongC., FischerL.A., and TheunissenT.W. (2019). Recent insights into the naive state of human pluripotency and its applications. Exp. Cell. Res. 385,111645.
9.
EstebanM.A., XuJ., YangJ., PengM., QinD., LiW., JiangZ., ChenJ., DengK., ZhongM., CaiJ., LaiL., and PeiD. (2009). Generation of induced pluripotent stem cell lines from Tibetan miniature pig. J. Biol. Chem. 284, 17634–17640.
10.
EzashiT., TeluguB.P., AlexenkoA.P., SachdevS., SinhaS., and RobertsR.M. (2009). Derivation of induced pluripotent stem cells from pig somatic cells. Proc. Natl. Acad. Sci. U. S. A. 106, 10993–10998.
FujishiroS.H., NakanoK., MizukamiY., AzamiT., AraiY., MatsunariH., IshinoR., NishimuraT., WatanabeM., AbeT., FurukawaY., UmeyamaK., YamanakaS., EmaM., NagashimaH., and HanazonoY. (2013). Generation of naive-like porcine-induced pluripotent stem cells capable of contributing to embryonic and fetal development. Stem Cells Dev. 22, 473–482.
13.
FukudaT., TaniT., HaraguchiS., DonaiK., NakajimaN., UenishiH., EitsukaT., MiyagawaM., SongS., OnumaM., HoshinoY., SatoE., and HondaA. (2017). Expression of six proteins causes reprogramming of porcine fibroblasts into induced pluripotent stem cells with both active X chromosomes. J. Cell Biochem. 118, 537–553.
GaoY., GuoY., DuanA., ChengD., ZhangS., and WangH. (2014). Optimization of culture conditions for maintaining porcine induced pluripotent stem cells. DNA Cell Biol. 33, 1–11.
16.
HaghighiF., DahlmannJ., Nakhaei-RadS., LangA., KutschkaI., ZenkerM., KensahG., PiekorzR. P., and AhmadianM. R. (2018). bFGF-mediated pluripotency maintenance in human induced pluripotent stem cells is associated with NRAS-MAPK signaling. Cell Commun. Signal. 16,96.
17.
HondaA., HatoriM., HiroseM., HondaC., IzuH., InoueK., HirasawaR., MatobaS., TogayachiS., MiyoshiH., and OguraA. (2013). Naive-like conversion overcomes the limited differentiation capacity of induced pluripotent stem cells. J. Biol. Chem. 288, 26157–26166.
18.
HouD.R., JinY., NieX.W., ZhangM.L., TaN., ZhaoL.H., YangN., ChenY., WuZ.Q., JiangH.B., LiY.R., SunQ.Y., DaiY.F., and LiR.F. (2016). Derivation of porcine embryonic stem-like cells from in vitro-produced blastocyst-stage embryos. Sci. Rep. 6,25838.
19.
KimE., KimM., HwangS.U., KimJ., LeeG., ParkY.S., and HyunS.H. (2019). Neural induction of porcine-induced pluripotent stem cells and further differentiation using glioblastoma-cultured medium. J. Cell. Mol. Med. 23, 2052–2063.
20.
LiD., SecherJ., HyttelP., IvaskM., KolkoM., HallV.J., and FreudeK.K. (2018). Generation of transgene-free porcine intermediate type induced pluripotent stem cells. Cell Cycle, 17, 2547–2563.
21.
LiuY., and YamashitaJ.K. (2019). AMPK activators contribute to maintain naive pluripotency in mouse embryonic stem cells. Biochem. Biophys. Res. Commun. 509, 24–31.
22.
LiuY., YangJ.Y., LuY., YuP., DoveC.R., HutchesonJ.M., MumawJ.L., SticeS.L., and WestF.D. (2013). alpha-1,3-Galactosyltransferase knockout pig induced pluripotent stem cells: A cell source for the production of xenotransplant pigs. Cell. Reprogram. 15, 107–116.
23.
NicholsJ., and SmithA. (2009). Naive and primed pluripotent states. Cell Stem Cell, 4, 487–492.
24.
ParkT.S., ZimmerlinL., Evans-MosesR., and ZambidisE.T. (2018a). Chemical reversion of conventional human pluripotent stem cells to a naïve-like state with improved multilineage differentiation potency. J. Vis. Exp. 136,57921.
25.
ParkT.Y., ChoiK.H., LeeD.K., OhJ.N., KimS.H., and LeeC.K. (2018b). Attempting to convert primed porcine embryonic stem cells into a naive state through the overexpression of reprogramming factors. Cell. Reprogram. 20, 289–300.
26.
QinM., ChenR., LiH., LiangH., XueQ., LiF., ChenY., and ZhangX. (2016). Direct reprogramming of human amniotic fluid stem cells by OCT4 and application in repairing of cerebral ischemia damage. Int. J. Biol. Sci. 12, 558–568.
27.
SetthawongP., PhakdeedindanP., TiptanavattanaN., RungarunlertS., TechakumphuM., and TharasanitT. (2019). Generation of porcine induced-pluripotent stem cells from Sertoli cells. Theriogenology, 127, 32–40.
28.
SuY., ZhuJ., SalmanS., and TangY. (2020). Induced pluripotent stem cells from farm animals. J. Anim. Sci. 98,skaa343.
29.
TakahashiK., and YamanakaS. (2006). Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell, 126, 663–676.
30.
TakahashiS., KobayashiS., and HirataniI. (2018). Epigenetic differences between naive and primed pluripotent stem cells. Cell Mol. Life Sci. 75, 1191–1203.
31.
TangL., YinY., ZhouH., SongG., FanA., TangB., ShiW., and LiZ. (2012). Proliferative capacity and pluripotent characteristics of porcine adult stem cells derived from adipose tissue and bone marrow. Cell. Reprogram. 14, 342–352.
32.
TatP.A., SumerH., JonesK.L., UptonK., and VermaP.J. (2010). The efficient generation of induced pluripotent stem (iPS) cells from adult mouse adipose tissue-derived and neural stem cells. Cell Transplant. 19, 525–536.
33.
TeluguB.P., EzashiT., and RobertsR.M. (2010). Porcine induced pluripotent stem cells analogous to naive and primed embryonic stem cells of the mouse. Int. J. Dev. Biol. 54, 1703–1711.
34.
WareC.B., NelsonA.M., MechamB., HessonJ., ZhouW., JonlinE.C., Jimenez-CalianiA.J., DengX., CavanaughC., CookS., TesarP.J., OkadaJ., MargarethaL., SperberH., ChoiM., BlauC.A., TreutingP.M., HawkinsR.D., CirulliV., and Ruohola-BakerH. (2014). Derivation of naive human embryonic stem cells. Proc. Natl. Acad. Sci. U. S. A. 111, 4484–4489.
35.
WuZ., ChenJ., RenJ., BaoL., LiaoJ., CuiC., RaoL., LiH., GuY., DaiH., ZhuH., TengX., ChengL., and XiaoL. (2009). Generation of pig induced pluripotent stem cells with a drug-inducible system. J. Mol. Cell. Biol. 1, 46–54.
36.
XuJ., YuL., GuoJ., XiangJ., ZhengZ., GaoD., ShiB., HaoH., JiaoD., ZhongL., WangY., WuJ., WeiH., and HanJ. (2019). Generation of pig induced pluripotent stem cells using an extended pluripotent stem cell culture system. Stem Cell Res. Ther. 10,193.
37.
YuanY., ParkJ., TianY., ChoiJ., and PasquarielloR. (2019). A six-inhibitor culture medium for improving naive-type pluripotency of porcine pluripotent stem cells. Cell Death Discov. 5,104.
38.
ZhangS., GuoY., CuiY., LiuY., YuT., and WangH. (2015a). Generation of intermediate porcine iPS cells under culture condition favorable for mesenchymal-to-epithelial transition. Stem Cell Rev. Rep. 11, 24–38.
39.
ZhangW., PeiY., ZhongL., WenB., CaoS., and HanJ. (2015b). Pluripotent and metabolic features of two types of porcine iPSCs derived from defined mouse and human ES cell culture conditions. PLoS One 10,e0124562.
40.
ZhangY., WeiC., ZhangP., LiX., LiuT., PuY., LiY., CaoZ., CaoH., LiuY., ZhangX., and ZhangY. (2014). Efficient reprogramming of naive-like induced pluripotent stem cells from porcine adipose-derived stem cells with a feeder-independent and serum-free system. PLoS One 9,e85089.
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.
