PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of f1000medLatest ContentReportsReportsReports
 
F1000 Med Rep. 2010; 2: 44.
Published online 2010 June 16. doi:  10.3410/M2-44
PMCID: PMC2950048

Reconstructing blood from induced pluripotent stem cells

Abstract

The direct reprogramming of human somatic cells to induced pluripotent stem cells (iPSCs) offers exciting prospects for disease modelling and regenerative medicine. Several recent reports support the feasibility of generating various blood cell types from iPSCs through in vitro-directed differentiation. However, the derivation of hematopoietic stem cells (HSCs) capable of long-term reconstitution of all hematopoietic lineages appears to be more challenging. These hurdles notwithstanding, cell engineering strategies aiming to correct genetic defects at the stem cell level are already emerging. Robust methodologies for the generation of definitive human HSCs conferring high-level, multilineage, long-term, hematopoietic reconstitution thus are direly needed before the therapeutic potential and safety of iPSC-derived cell products can be thoroughly investigated.

Introduction and context

In 2006, Takahashi and Yamanaka [1] reported the direct reprogramming of mouse fibroblasts to pluripotent stem cells, which were termed induced pluripotent stem cells (iPSCs). This scientific breakthrough was achieved through overexpression of the four transcription factors OCT4, SOX2, KLF4, and c-MYC [1]. The successful reprogramming of human somatic cells reported the following year [2-4] opened new frontiers for disease modelling and regenerative medicine. Since then, several methodologies, using viral, non-viral, or chemical approaches, have been devised to establish iPSCs from various mouse and human somatic cell types [5]. These include mouse cells in all stages of the hematopoietic hierarchy and human progenitor cells from cord blood, bone marrow, and peripheral blood [6-9]. It is now widely accepted that mouse and human iPSCs possess morphological, molecular, and developmental attributes that closely resemble those of blastocyst-derived embryonic stem cells (ESCs), although their global gene expression patterns and epigenetic state may not be identical [5,10]. Directed differentiation protocols already in use in ESCs were promptly applied to iPSCs, yielding numerous cell types, including neurons, cardiomyocytes, adipocytes, and endothelial and hematopoietic cells.

However, the generation of human hematopoietic stem cells (HSCs) conferring multilineage, long-term, hematopoietic reconstitution remains elusive. HSCs originate in early embryonic development, after which they expand and self-renew in successive anatomical locations. Hematopoiesis is initiated with the specification of a subset of mesodermal cells known as hemangioblasts, which yield both endothelial and hematopoietic progeny. The first hemangioblasts arise in the primitive streak of the embryo and then migrate to the yolk sac, where they form blood islands [11]. The ensuing first wave of hematopoiesis predominantly yields erythroid cells, which express embryonic and fetal globins and macrophages, but not lymphoid cells. The immediate precursors of definitive HSCs are arterial endothelial cells [12-15], which become HSCs capable of long-term multilineage repopulation of adult hosts in the dorsal aorta of the aorta-gonad-mesonephros (AGM) region and the chorioallantoic vessels of the placenta. These early CD34+, c-kit+, CD41+ HSCs next migrate to the placenta and mostly to the fetal liver, where their numbers further expand. Around birth, the main site of hematopoiesis shifts from liver to bone marrow, from which most blood cells originate throughout the organism’s adult life span.

Recent advances

In vitro generation of blood cell types from ESCs and iPSCs

Hematopoietic lineage specification from pluripotent cells is obtained in one of two general approaches, using either embryoid body formation in the presence of hematopoietic cytokines or co-culture with stromal cell lines such as the OP9 stroma cell line. Several blood cell types have been successfully generated from murine or human ESCs. These include mouse and human B- and T-lineage cells [16], megakaryocytes [17], and erythroid cells. The latter have revealed that human ESC-derived erythropoiesis closely mimics primitive erythropoiesis, characterized by the expression of the embryonic ε and ζ and the fetal α and γ globins [18,19]. The analysis of globin gene expression in erythrocytes derived from human iPSCs likewise revealed a pattern of primitive erythropoiesis [20].

Derivation of HSCs from ECSs and iPSCs

The generation of self-renewing multipotent HSCs from ESCs or iPSCs appears to be challenging. Several early reports illustrated the difficulty of producing HSCs capable of reconstituting adult, irradiated recipients [21,22]. Intravenous injection of murine or human ESC- and iPSC-derived hematopoietic cells resulted in little or no engraftment [23-28]. More recent studies have yielded more encouraging results. Ledran et al. [29] reported that co-culture of human ESCs on primary AGM stroma induced HSCs capable of primary and secondary hematopoietic engraftment into nonobese diabetic/severe combined immunodeficiency disease Il2rgcnull (NOG) mice (0.11-16.26%). Furthermore, culture of human ESCs and iPSCs with medium conditioned by HepG2 cells, a human hepatocarcinoma cell line, was shown to enhance the generation of mesodermal derivatives, including hematopoietic cells, with the ability to repopulate sublethally irradiated NOG mice for almost 1 year [30].

Efforts to engineer functional adult HSCs from ESCs or iPSCs

Attempts to engineer definitive hematopoiesis have so far largely focused on HOX family members (particularly HOXB4 and HOXA10), which are transcription factors involved in the formation or maintenance of HSCs [11]. Ectopic expression of HOXB4 in mouse ESCs has been reported to give rise to HSC-like cells capable of long-term repopulation [31,32]. However, the in vivo reconstitution obtained is strongly biased toward myeloid cells, hardly yielding lymphoid cells [32,33], while continuous HOXB4 expression poses oncogenic risks [34]. Cdx genes were also shown to promote the specification of hematopoietic progenitors in mouse ESCs [35]. However, these strategies have not been successful toward the generation of engraftable HSCs from human ESCs [25], highlighting the difficulty in translating mouse ESC-based studies into human pluripotent stem cell engineering.

Several investigators have observed that globin switching occurs in cultured human ESC-derived erythroid progeny in a time-dependent fashion, although the underlying mechanisms are not understood [18,19,36]. Fetal hematopoietic cells transplanted into adult sheep were previously shown to switch over time [37]. It is therefore possible that a prolonged in vitro culture or in vivo maturation is needed for the generation of developmentally mature erythroid progeny.

Implications for clinical practice

The advent of iPSCs holds great promise for regenerative medicine. iPSC-based research is poised to enable a watershed of knowledge on human cell development as well as provide critical tools for disease modelling and in vitro drug screening. The potential for developing novel cell therapies is equally tantalizing but still uncertain at this time. Reprogramming technologies offer the prospect of generating blood cell types on a patient-specific basis. Thus, red blood cells could be generated for individuals with severe congenital anemias or polytransfused subjects who require a unique blood cell type. A repertoire of naïve T lymphocytes could be generated for subjects with acquired or congenital deficiencies or aging subjects afflicted by immunosenescence. However, the methods for generating such cell types are still in their infancy and marred with considerable biological, safety, and economic uncertainties. A holy grail for this field is to generate HSCs, which would open new doors for both disease correction and regenerative medicine. Two recent proof-of-principle studies highlighted the potential of combined gene and cell therapy with autologous iPSCs to treat Fanconi anemia and sickle cell disease [26,33]. Robust protocols for the directed differentiation of human iPSCs to all hematopoietic cells will provide valuable tools for modelling hematopoiesis and hematological disorders, as exemplified in a recent study on polycythemia vera [9]. However, as reviewed above, there remain significant obstacles to the generation of adult HSCs capable of long-term, pan-lineage, hematopoietic reconstitution. Further studies are also required to assess the hematopoietic potential of human iPSCs in comparison with that of ESCs [9,38-40]. Thus, the well-deserved excitement about iPSCs must be tempered with patience as a considerably larger body of basic investigation is needed to unravel the biological and therapeutic potential as well as the safety profile of iPSC-derived hematopoietic cell products.

Acknowledgments

The authors are supported by grants from the Starr Foundation (Tri-Institutional Stem Cell Initiative, Tri-SCI-018), New York State Stem Cell Science (NYSTEM) (N08T-060) and the US National Institutes of Health (HL053750).

Abbreviations

AGM
aorta-gonad-mesonephros
ESC
embryonic stem cell
HSC
hematopoietic stem cell
iPSC
induced pluripotent stem cell
NOG
nonobese diabetic/severe combined immunodeficiency disease Il2rgcnull

Notes

The electronic version of this article is the complete one and can be found at: http://f1000.com/reports/m/2/44

Notes

Competing Interests

The authors declare that they have no competing interests.

References

1. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126:663–76. doi: 10.1016/j.cell.2006.07.024. [PubMed] [Cross Ref] F1000 Factor 10.5 Exceptional
Evaluated by Thomas Graf 30 Aug 2006, Wolf Reik 04 Sep 2006, Raphael Kopan 12 Sep 2006, Charles Coutelle 24 Oct 2006, Charles Brenner 11 Jun 2007
2. Takahashi K, Tanabe K, Ohnuki M, Narita M, Ichisaka T, Tomoda K, Yamanaka S. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131:861–72. doi: 10.1016/j.cell.2007.11.019. [PubMed] [Cross Ref] F1000 Factor 10.2 Exceptional
Evaluated by Thomas Graf 26 Nov 2007, Cheng-Ming Chiang 30 Nov 2007, John Mullins 03 Dec 2007, Maria Blasco 13 Dec 2007, Michael Cole 18 Dec 2007
3. Yu J, Vodyanik MA, Smuga-Otto K, Antosiewicz-Bourget J, Frane JL, Tian S, Nie J, Jonsdottir GA, Ruotti V, Stewart R, Slukvin II, Thomson JA. Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007;318:1917–20. doi: 10.1126/science.1151526. [PubMed] [Cross Ref] F1000 Factor 9.0 Exceptional
Evaluated by Cheng-Ming Chiang 30 Nov 2007
4. Park IH, Zhao R, West JA, Yabuuchi A, Huo H, Ince TA, Lerou PH, Lensch MW, Daley GQ. Reprogramming of human somatic cells to pluripotency with defined factors. Nature. 2008;451:141–6. doi: 10.1038/nature06534. [PubMed] [Cross Ref]
5. Kiskinis E, Eggan K. Progress toward the clinical application of patient-specific pluripotent stem cells. J Clin Invest. 2010;120:51–9. doi: 10.1172/JCI40553. [PMC free article] [PubMed] [Cross Ref]
6. Giorgetti A, Montserrat N, Aasen T, Gonzalez F, Rodríguez-Pizà I, Vassena R, Raya A, Boué S, Barrero MJ, Corbella BA, Torrabadella M, Veiga A, Izpisua Belmonte JC. Generation of induced pluripotent stem cells from human cord blood using OCT4 and SOX2. Cell Stem Cell. 2009;5:353–7. doi: 10.1016/j.stem.2009.09.008. [PMC free article] [PubMed] [Cross Ref]
7. Haase A, Olmer R, Schwanke K, Wunderlich S, Merkert S, Hess C, Zweigerdt R, Gruh I, Meyer J, Wagner S, Maier LS, Han DW, Glage S, Miller K, Fischer P, Schöler HR, Martin U. Generation of induced pluripotent stem cells from human cord blood. Cell Stem Cell. 2009;5:434–41. doi: 10.1016/j.stem.2009.08.021. [PubMed] [Cross Ref]
8. Loh YH, Agarwal S, Park IH, Urbach A, Huo H, Heffner GC, Kim K, Miller JD, Ng K, Daley GQ. Generation of induced pluripotent stem cells from human blood. Blood. 2009;113:5476–9. doi: 10.1182/blood-2009-02-204800. [PubMed] [Cross Ref] F1000 Factor 4.8 Must Read
Evaluated by Anthony D Ho 01 Jun 2009, James Ellis 01 Apr 2009
9. Ye Z, Zhan H, Mali P, Dowey S, Williams DM, Jang YY, Dang CV, Spivak JL, Moliterno AR, Cheng L. Human-induced pluripotent stem cells from blood cells of healthy donors and patients with acquired blood disorders. Blood. 2009;114:5473–80. doi: 10.1182/blood-2009-04-217406. [PubMed] [Cross Ref] F1000 Factor 3.0 Recommended
Evaluated by Gay Crooks 20 Apr 2010
10. Chin MH, Mason MJ, Xie W, Volinia S, Singer M, Peterson C, Ambartsumyan G, Aimiuwu O, Richter L, Zhang J, Khvorostov I, Ott V, Grunstein M, Lavon N, Benvenisty N, Croce CM, Clark AT, Baxter T, Pyle AD, Teitell MA, Pelegrini M, Plath K, Lowry WE. Induced pluripotent stem cells and embryonic stem cells are distinguished by gene expression signatures. Cell Stem Cell. 2009;5:111–23. doi: 10.1016/j.stem.2009.06.008. [PubMed] [Cross Ref]
11. Orkin SH, Zon LI. Hematopoiesis: an evolving paradigm for stem cell biology. Cell. 2008;132:631–44. doi: 10.1016/j.cell.2008.01.025. [PMC free article] [PubMed] [Cross Ref]
12. Robert-Moreno A, Guiu J, Ruiz-Herguido C, López ME, Inglés-Esteve J, Riera L, Tipping A, Enver T, Dzierzak E, Gridley T, Espinosa L, Bigas A. Impaired embryonic haematopoiesis yet normal arterial development in the absence of the Notch ligand Jagged1. Embo J. 2008;27:1886–95. doi: 10.1038/emboj.2008.113. [PubMed] [Cross Ref]
13. Eilken HM, Nishikawa S, Schroeder T. Continuous single-cell imaging of blood generation from haemogenic endothelium. Nature. 2009;457:896–900. doi: 10.1038/nature07760. [PubMed] [Cross Ref]
14. Zovein AC, Hofmann JJ, Lynch M, French WJ, Turlo KA, Yang Y, Becker MS, Zanetta L, Dejana E, Gasson JC, Tallquist MD, Iruela-Arispe ML. Fate tracing reveals the endothelial origin of hematopoietic stem cells. Cell Stem Cell. 2008;3:625–36. doi: 10.1016/j.stem.2008.09.018. [PMC free article] [PubMed] [Cross Ref] F1000 Factor 6.0 Must Read
Evaluated by Marella De Bruijn 16 Feb 2009
15. Rhodes KE, Gekas C, Wang Y, Lux CT, Francis CS, Chan DN, Conway S, Orkin SH, Yoder MC, Mikkola HK. The emergence of hematopoietic stem cells is initiated in the placental vasculature in the absence of circulation. Cell Stem Cell. 2008;2:252–63. doi: 10.1016/j.stem.2008.01.001. [PMC free article] [PubMed] [Cross Ref] F1000 Factor 9.0 Exceptional
Evaluated by Hans-Willem Snoeck 19 Mar 2008
16. Timmermans F, Velghe I, Vanwalleghem L, De Smedt M, Van Coppernolle S, Taghon T, Moore HD, Leclercq G, Langerak AW, Kerre T, Plum J, Vandekerckhove B. Generation of T cells from human embryonic stem cell-derived hematopoietic zones. J Immunol. 2009;182:6879–88. doi: 10.4049/jimmunol.0803670. [PubMed] [Cross Ref] F1000 Factor 9.0 Exceptional
Evaluated by Astar Winoto 08 Jun 2009
17. Takayama N, Nishikii H, Usui J, Tsukui H, Sawaguchi A, Hiroyama T, Eto K, Nakauchi H. Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors. Blood. 2008;111:5298–306. doi: 10.1182/blood-2007-10-117622. [PubMed] [Cross Ref]
18. Lu SJ, Feng Q, Park JS, Vida L, Lee BS, Strausbauch M, Wettstein PJ, Honig GR, Lanza R. Biologic properties and enucleation of red blood cells from human embryonic stem cells. Blood. 2008;112:4475–84. doi: 10.1182/blood-2008-05-157198. [PubMed] [Cross Ref] F1000 Factor 3.0 Recommended
Evaluated by Thalia Papayannopoulou 09 Sep 2008
19. Qiu C, Olivier EN, Velho M, Bouhassira EE. Globin switches in yolk sac-like primitive and fetal-like definitive red blood cells produced from human embryonic stem cells. Blood. 2008;111:2400–8. doi: 10.1182/blood-2007-07-102087. [PubMed] [Cross Ref]
20. Chang KH, Huang A, Hirata RK, Wang PR, Russell DW, Papayannopoulou T. Globin phenotype of erythroid cells derived from human induced pluripotent stem cells. Blood. 115:2553–4. doi: 10.1182/blood-2009-11-252650. [PubMed] [Cross Ref]
21. Wang L, Menendez P, Cerdan C, Bhatia M. Hematopoietic development from human embryonic stem cell lines. Exp Hematol. 2005;33:987–96. doi: 10.1016/j.exphem.2005.06.002. [PubMed] [Cross Ref]
22. Lengerke C, Daley GQ. Patterning definitive hematopoietic stem cells from embryonic stem cells. Exp Hematol. 2005;33:971–9. doi: 10.1016/j.exphem.2005.06.004. [PubMed] [Cross Ref]
23. Narayan AD, Chase JL, Lewis RL, Tian X, Kaufman DS, Thomson JA, Zanjani ED. Human embryonic stem cell-derived hematopoietic cells are capable of engrafting primary as well as secondary fetal sheep recipients. Blood. 2006;107:2180–3. doi: 10.1182/blood-2005-05-1922. [PubMed] [Cross Ref]
24. Lu SJ, Feng Q, Caballero S, Chen Y, Moore MA, Grant MB, Lanza R. Generation of functional hemangioblasts from human embryonic stem cells. Nat Methods. 2007;4:501–9. doi: 10.1038/nmeth1041. [PubMed] [Cross Ref]
25. Wang L, Menendez P, Shojaei F, Li L, Mazurier F, Dick JE, Cerdan C, Levac K, Bhatia M. Generation of hematopoietic repopulating cells from human embryonic stem cells independent of ectopic HOXB4 expression. J Exp Med. 2005;201:1603–14. doi: 10.1084/jem.20041888. [PMC free article] [PubMed] [Cross Ref]
26. Raya A, Rodríguez-Pizà I, Guenechea G, Vassena R, Navarro S, Barrero MJ, Consiglio A, Castellà M, Río P, Sleep E, González F, Tiscornia G, Garreta E, Aasen T, Veiga A, Verma IM, Surrallés J, Bueren J, Izpisúa Belmonte JC. Disease-corrected haematopoietic progenitors from Fanconi anaemia induced pluripotent stem cells. Nature. 2009;460:53–9. doi: 10.1038/nature08129. [PMC free article] [PubMed] [Cross Ref] F1000 Factor 9.8 Exceptional
Evaluated by James Ellis 15 Jun 2009, Laura Haneline 25 Jun 2009, Michel Sadelain 07 Sep 2009
27. Tian X, Woll PS, Morris JK, Linehan JL, Kaufman DS. Hematopoietic engraftment of human embryonic stem cell-derived cells is regulated by recipient innate immunity. Stem Cells. 2006;24:1370–80. doi: 10.1634/stemcells.2005-0340. [PubMed] [Cross Ref]
28. Tian X, Hexum MK, Penchev VR, Taylor RJ, Shultz LD, Kaufman DS. Bioluminescent imaging demonstrates that transplanted human embryonic stem cell-derived CD34(+) cells preferentially develop into endothelial cells. Stem Cells. 2009;27:2675–85. doi: 10.1002/stem.204. [PMC free article] [PubMed] [Cross Ref]
29. Ledran MH, Krassowska A, Armstrong L, Dimmick I, Renström J, Lang R, Yung S, Santibanez-Coref M, Dzierzak E, Stojkovic M, Oostendorp RA, Forrester L, Lako M. Efficient hematopoietic differentiation of human embryonic stem cells on stromal cells derived from hematopoietic niches. Cell Stem Cell. 2008;3:85–98. doi: 10.1016/j.stem.2008.06.001. [PubMed] [Cross Ref] F1000 Factor 3.0 Recommended
Evaluated by Dan Kaufman 29 Jul 2008
30. Lu M, Kardel MD, O’Connor MD, Eaves CJ. Enhanced generation of hematopoietic cells from human hepatocarcinoma cell-stimulated human embryonic and induced pluripotent stem cells. Exp Hematol. 2009;37:924–36. doi: 10.1016/j.exphem.2009.05.007. [PubMed] [Cross Ref]
31. Kyba M, Perlingeiro RC, Daley GQ. HoxB4 confers definitive lymphoid-myeloid engraftment potential on embryonic stem cell and yolk sac hematopoietic progenitors. Cell. 2002;109:29–37. doi: 10.1016/S0092-8674(02)00680-3. [PubMed] [Cross Ref] F1000 Factor 6.0 Must Read
Evaluated by Ana Cumano 22 May 2002
32. Wang Y, Yates F, Naveiras O, Ernst P, Daley GQ. Embryonic stem cell-derived hematopoietic stem cells. Proc Natl Acad Sci U S A. 2005;102:19081–6. doi: 10.1073/pnas.0506127102. [PubMed] [Cross Ref]
33. Hanna J, Wernig M, Markoulaki S, Sun CW, Meissner A, Cassady JP, Beard C, Brambrink T, Wu LC, Townes TM, Jaenisch R. Treatment of sickle cell anemia mouse model with iPS cells generated from autologous skin. Science. 2007;318:1920–3. doi: 10.1126/science.1152092. [PubMed] [Cross Ref] F1000 Factor 9.8 Exceptional
Evaluated by William H Colledge 04 Jan 2008, Fritz Eckstein 11 Jan 2008, Ali H Brivanlou 30 Jan 2008
34. Zhang XB, Beard BC, Trobridge GD, Wood BL, Sale GE, Sud R, Humphries RK, Kiem HP. High incidence of leukemia in large animals after stem cell gene therapy with a HOXB4-expressing retroviral vector. J Clin Invest. 2008;118:1502–10. doi: 10.1172/JCI34371. [PMC free article] [PubMed] [Cross Ref] F1000 Factor 3.0 Recommended
Evaluated by Kateri Moore 07 May 2008
35. McKinney-Freeman SL, Lengerke C, Jang IH, Schmitt S, Wang Y, Philitas M, Shea J, Daley GQ. Modulation of murine embryonic stem cell-derived CD41+c-kit+ hematopoietic progenitors by ectopic expression of Cdx genes. Blood. 2008;111:4944–53. doi: 10.1182/blood-2007-11-124644. [PubMed] [Cross Ref]
36. Ma F, Ebihara Y, Umeda K, Sakai H, Hanada S, Zhang H, Zaike Y, Tsuchida E, Nakahata T, Nakauchi H, Tsuji K. Generation of functional erythrocytes from human embryonic stem cell-derived definitive hematopoiesis. Proc Natl Acad Sci U S A. 2008;105:13087–92. doi: 10.1073/pnas.0802220105. [PubMed] [Cross Ref]
37. Wood WG, Bunch C. Fetal-to-adult hemopoietic cell transplantation: is hemoglobin synthesis gestational age-dependent? Prog Clin Biol Res. 1983;134:511–21. [PubMed]
38. Feng Q, Lu SJ, Klimanskaya I, Gomes I, Kim D, Chung Y, Honig GR, Kim KS, Lanza R. Hemangioblastic derivatives from human induced pluripotent stem cells exhibit limited expansion and early senescence. Stem Cells. 2010;28:704–12. [PubMed] F1000 Factor 3.0 Recommended
Evaluated by Gay Crooks 22 April 2010
39. Choi KD, Yu J, Smuga-Otto K, Salvagiotto G, Rehrauer W, Vodyanik M, Thomson J, Slukvin I. Hematopoietic and endothelial differentiation of human induced pluripotent stem cells. Stem Cells. 2009;27:559–67. doi: 10.1002/stem.20080922. [PMC free article] [PubMed] [Cross Ref]
40. Lengerke C, Grauer M, Niebuhr NI, Riedt T, Kanz L, Park IH, Daley GQ. Hematopoietic development from human induced pluripotent stem cells. Ann N Y Acad Sci. 2009;1176:219–27. doi: 10.1111/j.1749-6632.2009.04606.x. [PMC free article] [PubMed] [Cross Ref]

Articles from F1000 Medicine Reports are provided here courtesy of Faculty of 1000 Ltd