Stem Cell Engineering and Analysis
The hESC Core Facility is a basic and translational stem cell research facility situated in the Boston Children’s Hospital Stem Cell Program (Karp Building, RB09213
, 1 Blackfan Cirlce, Boston MA 02115). Areas of expertise include human pluripotent stem cell biology, cGMP cell manufacturing, reprogramming, genome editing, genotyping, laboratory automation, chemical screening, and imaging/image analysis. Our goal is to leverage stem cell technologies to gain novel insights into hematopoietic diseases and to accelerate the development of therapies.
PI/Director:
Thorsten Schlaeger, Ph.D.
617.919.2082
Current Stem Cell Core (Schlaeger lab) Members
Ana Capi
Arunoday Bhan
Audrey Tran
Brett Estes
Mike Shi
Nate Mullin
Yang Tang
Yosra Zhang
To request hiPSCs please contact
Aimee Dixon (Daley lab) or Thorsten Schlaeger (Core). See table for other services and fees for non-profit users (U54 members will routinely receive a 33% discount).
| Reprogramming Service | ||||
| Somatic Cell Material | 1.a. Isolation, expansion, and banking of skin biopsy fibroblasts | $1,000 | ||
| •• OR •• | ||||
| 1.b. Expansion and banking of frozen fibroblasts | $500 | |||
| •• OR •• | ||||
| 1.c. Isolation/banking/expansion of PB mononuclear cells | $750 | |||
| •• OR •• | ||||
| 1.d. Expansion of frozen mononuclear cells | $500 | |||
| 2. Somatic cell mycoplasma testing | $50 | |||
| 3. Somatic cell DNA isolation and fingerprinting | $300 | |||
| Somatic cell module total | $850-$1,350 | |||
| Reprogr. | 4a. SeV reprogramming (fibroblasts or PBMCs) | $2,000 | ||
| •• OR •• | ||||
| 4b. Epsiomal reprogramming (PBMCs) | $1,000 | |||
| Reprogramming total | $1,000-$2,000 | |||
| Banking and QC | 5. Clone picking, expansion, banking (P8, 5 vials, 3 clones) | $1,500 | ||
| 6. Immunofluorescence (OCT4, TRA160, CD90, NANOG, TRA181, SSEA5, E-Cadherin, SSEA4, Epcam, SSEA1),P10, 3 clones | $750 | |||
| 7. Karyotyping, P10, 3 clones | $2,100 | |||
| 8. DNA isolation and DNA fingerprinting, P10, 3 clones | $900 | |||
| 9. Mycoplasma testing, P10, 3 clones | $150 | |||
| 10. EBNA QPCR (episomal reprogramming), P10, 3 clones | $500 | |||
| Expansion, banking, and QC total | $5,400-$5,900 | |||
| Reprogramming project total | $7,750-$9,250 | |||
| Genome Editing Service | ||||
| hPSC Material | 1. Thaw, mycoplasma-test, expansion, adaptation to single-cell passaging | $1,000 | ||
| 2. Karyotyping (optional if recent karyotype is provided) | $1,000 | |||
| 3. DNA isolation and fingerprinting (optional if DNA fingerprint is provided) | $300 | |||
| 4. Targeting vector design, construction, sequencing | $5,000 | |||
| 5. gRNA plasmid design, construction, sequencing | $500 | |||
| 6. PCR screening assay development (if not provided) | $1,200 | |||
| 7. hPSC Cas9(WT)-GFP + gRNA + TV nucleofection, FACS sorting, single cell plating, clone picking/expansion | $6,000 | |||
| 8. PCR screening of >100 clones | $2,000 | |||
| 9. Expansion, QC (IF, FP, KT), and banking, per clone | $2,000 | |||
| Genome Editing Project Total (3 clones) | $14,000-$23,000 | |||
FACS Core: please contact Ron Mathieu
Alumni
Amelia Cianci, Andrew Ettenger, Blair Bell, Christian Skorik, Dipti Gupta, Gloria Lee, Holly Young, Iris Huo, Jade (Tahni) McPherson, Justine Miller, Karrie Chan, Kelly Fitzgerald, Leah Boyer, Mam (Sutheera) Ratanasirintrawoot, Manav Gupta, Michael Chen, Michelle Godfrey, Odelya Hartung, Phil Manos, Prerana Malwadkar, Sascha (Alex) Devine, Sumon Datta, Will Yee, Xiao Guan.
Publications
1: Doulatov S, Vo LT, Macari ER, Wahlster L, Kinney MA, Taylor AM, Barragan J, Gupta M, McGrath K, Lee HY, Humphries JM, DeVine A, Narla A, Alter BP, Beggs AH, Agarwal S, Ebert BL, Gazda HT, Lodish HF, Sieff CA, Schlaeger TM, Zon LI, Daley GQ. Drug discovery for Diamond-Blackfan anemia using reprogrammed hematopoietic progenitors. Sci Transl Med. 2017 Feb 8;9(376). pii: eaah5645. doi: 10.1126/scitranslmed.aah5645. PubMed PMID: 28179501.
2: Cortes M, Chen MJ, Stachura DL, Liu SY, Kwan W, Wright F, Vo LT, Theodore LN, Esain V, Frost IM, Schlaeger TM, Goessling W, Daley GQ, North TE. Developmental Vitamin D Availability Impacts Hematopoietic Stem Cell Production. Cell Rep. 2016 Oct 4;17(2):458-468. doi: 10.1016/j.celrep.2016.09.012. PubMed PMID: 27705794; PubMed Central PMCID: PMC5338633.
3: Li P, Lahvic JL, Binder V, Pugach EK, Riley EB, Tamplin OJ, Panigrahy D, Bowman TV, Barrett FG, Heffner GC, McKinney-Freeman S, Schlaeger TM, Daley GQ, Zeldin DC, Zon LI. Epoxyeicosatrienoic acids enhance embryonic haematopoiesis and adult marrow engraftment. Nature. 2015 Jul 23;523(7561):468-71. doi: 10.1038/nature14569. PubMed PMID: 26201599; PubMed Central PMCID: PMC4754787.
4: Rissone A, Weinacht KG, la Marca G, Bishop K, Giocaliere E, Jagadeesh J, Felgentreff K, Dobbs K, Al-Herz W, Jones M, Chandrasekharappa S, Kirby M, Wincovitch S, Simon KL, Itan Y, DeVine A, Schlaeger T, Schambach A, Sood R, Notarangelo LD, Candotti F. Reticular dysgenesis-associated AK2 protects hematopoietic stem and progenitor cell development from oxidative stress. J Exp Med. 2015 Jul 27;212(8):1185-202. doi: 10.1084/jem.20141286. Epub 2015 Jul 6. PubMed PMID: 26150473; PubMed Central PMCID: PMC4516804.
5: Kim PG, Nakano H, Das PP, Chen MJ, Rowe RG, Chou SS, Ross SJ, Sakamoto KM, Zon LI, Schlaeger TM, Orkin SH, Nakano A, Daley GQ. Flow-induced protein kinase A-CREB pathway acts via BMP signaling to promote HSC emergence. J Exp Med. 2015 May 4;212(5):633-48. doi: 10.1084/jem.20141514. Epub 2015 Apr 13. PubMed PMID: 25870201; PubMed Central PMCID: PMC4419355.
6: Jing L, Tamplin OJ, Chen MJ, Deng Q, Patterson S, Kim PG, Durand EM, McNeil A, Green JM, Matsuura S, Ablain J, Brandt MK, Schlaeger TM, Huttenlocher A, Daley GQ, Ravid K, Zon LI. Adenosine signaling promotes hematopoietic stem and progenitor cell emergence. J Exp Med. 2015 May 4;212(5):649-63. doi: 10.1084/jem.20141528. Epub 2015 Apr 13. PubMed PMID: 25870200; PubMed Central PMCID: PMC4419349.
7: Jang IH, Lu YF, Zhao L, Wenzel PL, Kume T, Datta SM, Arora N, Guiu J, Lagha M, Kim PG, Do EK, Kim JH, Schlaeger TM, Zon LI, Bigas A, Burns CE, Daley GQ. Notch1 acts via Foxc2 to promote definitive hematopoiesis via effects on hemogenic endothelium. Blood. 2015 Feb 26;125(9):1418-26. doi: 10.1182/blood-2014-04-568170. Epub 2015 Jan 13. PubMed PMID: 25587036; PubMed Central PMCID: PMC4342355.
8: Schlaeger TM, Daheron L, Brickler TR, Entwisle S, Chan K, Cianci A, DeVine A, Ettenger A, Fitzgerald K, Godfrey M, Gupta D, McPherson J, Malwadkar P, Gupta M, Bell B, Doi A, Jung N, Li X, Lynes MS, Brookes E, Cherry AB, Demirbas D, Tsankov AM, Zon LI, Rubin LL, Feinberg AP, Meissner A, Cowan CA, Daley GQ. A comparison of non-integrating reprogramming methods. Nat Biotechnol. 2015 Jan;33(1):58-63. doi: 10.1038/nbt.3070. Epub 2014 Dec 1. PubMed PMID: 25437882; PubMed Central PMCID: PMC4329913.
9: Yien YY, Robledo RF, Schultz IJ, Takahashi-Makise N, Gwynn B, Bauer DE, Dass A, Yi G, Li L, Hildick-Smith GJ, Cooney JD, Pierce EL, Mohler K, Dailey TA, Miyata N, Kingsley PD, Garone C, Hattangadi SM, Huang H, Chen W, Keenan EM, Shah DI, Schlaeger TM, DiMauro S, Orkin SH, Cantor AB, Palis J, Koehler CM, Lodish HF, Kaplan J, Ward DM, Dailey HA, Phillips JD, Peters LL, Paw BH. TMEM14C is required for erythroid mitochondrial heme metabolism. J Clin Invest. 2014 Oct;124(10):4294-304. doi: 10.1172/JCI76979. Epub 2014 Aug 26. PubMed PMID: 25157825; PubMed Central PMCID: PMC4191016.
10: Sinkevicius KW, Kriegel C, Bellaria KJ, Lee J, Lau AN, Leeman KT, Zhou P, Beede AM, Fillmore CM, Caswell D, Barrios J, Wong KK, Sholl LM, Schlaeger TM, Bronson RT, Chirieac LR, Winslow MM, Haigis MC, Kim CF. Neurotrophin receptor TrkB promotes lung adenocarcinoma metastasis. Proc Natl Acad Sci U S A. 2014 Jul 15;111(28):10299-304. doi: 10.1073/pnas.1404399111. Epub 2014 Jun 30. PubMed PMID: 24982195; PubMed Central PMCID: PMC4104911.
11: Felgentreff K, Du L, Weinacht KG, Dobbs K, Bartish M, Giliani S, Schlaeger T, DeVine A, Schambach A, Woodbine LJ, Davies G, Baxi SN, van der Burg M, Bleesing J, Gennery A, Manis J, Pan-Hammarström Q, Notarangelo LD. Differential role of nonhomologous end joining factors in the generation, DNA damage response, and myeloid differentiation of human induced pluripotent stem cells. Proc Natl Acad Sci U S A. 2014 Jun 17;111(24):8889-94. doi: 10.1073/pnas.1323649111. Epub 2014 Jun 2. PubMed PMID: 24889605; PubMed Central PMCID: PMC4066476.
12: Chung J, Anderson SA, Gwynn B, Deck KM, Chen MJ, Langer NB, Shaw GC, Huston NC, Boyer LF, Datta S, Paradkar PN, Li L, Wei Z, Lambert AJ, Sahr K, Wittig JG, Chen W, Lu W, Galy B, Schlaeger TM, Hentze MW, Ward DM, Kaplan J, Eisenstein RS, Peters LL, Paw BH. Iron regulatory protein-1 protects against mitoferrin-1-deficient porphyria. J Biol Chem. 2014 Mar 14;289(11):7835-43. doi: 10.1074/jbc.M114.547778. Epub 2014 Feb 7. Erratum in: J Biol Chem. 2014 May 16;289(20):13707. PubMed PMID: 24509859; PubMed Central PMCID: PMC3953295.
13: Prilutsky D, Palmer NP, Smedemark-Margulies N, Schlaeger TM, Margulies DM, Kohane IS. iPSC-derived neurons as a higher-throughput readout for autism: promises and pitfalls. Trends Mol Med. 2014 Feb;20(2):91-104. doi: 10.1016/j.molmed.2013.11.004. Epub 2013 Dec 24. Review. PubMed PMID: 24374161; PubMed Central PMCID: PMC4117413.
14: Tulpule A, Kelley JM, Lensch MW, McPherson J, Park IH, Hartung O, Nakamura T, Schlaeger TM, Shimamura A, Daley GQ. Pluripotent stem cell models of Shwachman-Diamond syndrome reveal a common mechanism for pancreatic and hematopoietic dysfunction. Cell Stem Cell. 2013 Jun 6;12(6):727-36. doi: 10.1016/j.stem.2013.04.002. Epub 2013 Apr 18. PubMed PMID: 23602541; PubMed Central PMCID: PMC3755012.
15: Lafaille FG, Pessach IM, Zhang SY, Ciancanelli MJ, Herman M, Abhyankar A, Ying SW, Keros S, Goldstein PA, Mostoslavsky G, Ordovas-Montanes J, Jouanguy E, Plancoulaine S, Tu E, Elkabetz Y, Al-Muhsen S, Tardieu M, Schlaeger TM, Daley GQ, Abel L, Casanova JL, Studer L, Notarangelo LD. Impaired intrinsic immunity to HSV-1 in human iPSC-derived TLR3-deficient CNS cells. Nature. 2012 Nov 29;491(7426):769-73. doi: 10.1038/nature11583. Epub 2012 Oct 28. PubMed PMID: 23103873; PubMed Central PMCID: PMC3527075.
16: Müller LU, Schlaeger TM, DeVine AL, Williams DA. Induced pluripotent stem cells as a tool for gaining new insights into Fanconi anemia. Cell Cycle. 2012 Aug 15;11(16):2985-90. doi: 10.4161/cc.21109. Epub 2012 Jul 24. PubMed PMID: 22825249; PubMed Central PMCID: PMC3442908.
17: Müller LU, Milsom MD, Harris CE, Vyas R, Brumme KM, Parmar K, Moreau LA, Schambach A, Park IH, London WB, Strait K, Schlaeger T, Devine AL, Grassman E, D'Andrea A, Daley GQ, Williams DA. Overcoming reprogramming resistance of Fanconi anemia cells. Blood. 2012 Jun 7;119(23):5449-57. doi: 10.1182/blood-2012-02-408674. Epub 2012 Feb 27. PubMed PMID: 22371882; PubMed Central PMCID: PMC3369681.
18: Hyde BB, Liesa M, Elorza AA, Qiu W, Haigh SE, Richey L, Mikkola HK, Schlaeger TM, Shirihai OS. The mitochondrial transporter ABC-me (ABCB10), a downstream target of GATA-1, is essential for erythropoiesis in vivo. Cell Death Differ. 2012 Jul;19(7):1117-26. doi: 10.1038/cdd.2011.195. Epub 2012 Jan 13. PubMed PMID: 22240895; PubMed Central PMCID: PMC3374075.
19: Manos PD, Ratanasirintrawoot S, Loewer S, Daley GQ, Schlaeger TM. Live-cell immunofluorescence staining of human pluripotent stem cells. Curr Protoc Stem Cell Biol. 2011 Dec;Chapter 1:Unit 1C.12. doi: 10.1002/9780470151808.sc01c12s19. PubMed PMID: 22135082.
20: Miller JD, Schlaeger TM. Generation of induced pluripotent stem cell lines from human fibroblasts via retroviral gene transfer. Methods Mol Biol. 2011;767:55-65. doi: 10.1007/978-1-61779-201-4_5. PubMed PMID: 21822867.
21: Goessling W, Allen RS, Guan X, Jin P, Uchida N, Dovey M, Harris JM, Metzger ME, Bonifacino AC, Stroncek D, Stegner J, Armant M, Schlaeger T, Tisdale JF, Zon LI, Donahue RE, North TE. Prostaglandin E2 enhances human cord blood stem cell xenotransplants and shows long-term safety in preclinical nonhuman primate transplant models. Cell Stem Cell. 2011 Apr 8;8(4):445-58. doi: 10.1016/j.stem.2011.02.003. PubMed PMID: 21474107; PubMed Central PMCID: PMC3148081.
22: Luong MX, Auerbach J, Crook JM, Daheron L, Hei D, Lomax G, Loring JF, Ludwig T, Schlaeger TM, Smith KP, Stacey G, Xu RH, Zeng F. A call for standardized naming and reporting of human ESC and iPSC lines. Cell Stem Cell. 2011 Apr 8;8(4):357-9. doi: 10.1016/j.stem.2011.03.002. PubMed PMID: 21474098.
23: Pessach IM, Ordovas-Montanes J, Zhang SY, Casanova JL, Giliani S, Gennery AR, Al-Herz W, Manos PD, Schlaeger TM, Park IH, Rucci F, Agarwal S, Mostoslavsky G, Daley GQ, Notarangelo LD. Induced pluripotent stem cells: a novel frontier in the study of human primary immunodeficiencies. J Allergy Clin Immunol. 2011 Jun;127(6):1400-7.e4. doi: 10.1016/j.jaci.2010.11.008. Epub 2010 Dec 24. PubMed PMID: 21185069; PubMed Central PMCID: PMC3081993.
24: Loewer S, Cabili MN, Guttman M, Loh YH, Thomas K, Park IH, Garber M, Curran M, Onder T, Agarwal S, Manos PD, Datta S, Lander ES, Schlaeger TM, Daley GQ, Rinn JL. Large intergenic non-coding RNA-RoR modulates reprogramming of human induced pluripotent stem cells. Nat Genet. 2010 Dec;42(12):1113-7. doi: 10.1038/ng.710. Epub 2010 Nov 7. Erratum in: Nat Genet. 2010 Dec;42(12): 3 p following 1117. PubMed PMID: 21057500; PubMed Central PMCID: PMC3040650.
25: Warren L, Manos PD, Ahfeldt T, Loh YH, Li H, Lau F, Ebina W, Mandal PK, Smith ZD, Meissner A, Daley GQ, Brack AS, Collins JJ, Cowan C, Schlaeger TM, Rossi DJ. Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell. 2010 Nov 5;7(5):618-30. doi: 10.1016/j.stem.2010.08.012. Epub 2010 Sep 30. PubMed PMID: 20888316; PubMed Central PMCID: PMC3656821.
26: Hartung O, Huo H, Daley GQ, Schlaeger TM. Clump passaging and expansion of human embryonic and induced pluripotent stem cells on mouse embryonic fibroblast feeder cells. Curr Protoc Stem Cell Biol. 2010 Aug;Chapter 1:Unit 1C.10. doi: 10.1002/9780470151808.sc01c10s14. PubMed PMID: 20814935.
27: Tulpule A, Lensch MW, Miller JD, Austin K, D'Andrea A, Schlaeger TM, Shimamura A, Daley GQ. Knockdown of Fanconi anemia genes in human embryonic stem cells reveals early developmental defects in the hematopoietic lineage. Blood. 2010 Apr 29;115(17):3453-62. doi: 10.1182/blood-2009-10-246694. Epub 2010 Jan 20. PubMed PMID: 20089964; PubMed Central PMCID: PMC2867260.
28: Doi A, Park IH, Wen B, Murakami P, Aryee MJ, Irizarry R, Herb B, Ladd-Acosta C, Rho J, Loewer S, Miller J, Schlaeger T, Daley GQ, Feinberg AP. Differential methylation of tissue- and cancer-specific CpG island shores distinguishes human induced pluripotent stem cells, embryonic stem cells and fibroblasts. Nat Genet. 2009 Dec;41(12):1350-3. doi: 10.1038/ng.471. Epub 2009 Nov 1. PubMed PMID: 19881528; PubMed Central PMCID: PMC2958040.
29: Chan EM, Ratanasirintrawoot S, Park IH, Manos PD, Loh YH, Huo H, Miller JD, Hartung O, Rho J, Ince TA, Daley GQ, Schlaeger TM. Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells. Nat Biotechnol. 2009 Nov;27(11):1033-7. doi: 10.1038/nbt.1580. Epub 2009 Oct 11. PubMed PMID: 19826408.
30: Ficarro SB, Zhang Y, Lu Y, Moghimi AR, Askenazi M, Hyatt E, Smith ED, Boyer L, Schlaeger TM, Luckey CJ, Marto JA. Improved electrospray ionization efficiency compensates for diminished chromatographic resolution and enables proteomics analysis of tyrosine signaling in embryonic stem cells. Anal Chem. 2009 May 1;81(9):3440-7. doi: 10.1021/ac802720e. PubMed PMID: 19331382.
31: Lerou PH, Yabuuchi A, Huo H, Miller JD, Boyer LF, Schlaeger TM, Daley GQ. Derivation and maintenance of human embryonic stem cells from poor-quality in vitro fertilization embryos. Nat Protoc. 2008;3(5):923-33. doi: 10.1038/nprot.2008.60. PubMed PMID: 18451800.
32: Lensch MW, Schlaeger TM, Zon LI, Daley GQ. Teratoma formation assays with human embryonic stem cells: a rationale for one type of human-animal chimera. Cell Stem Cell. 2007 Sep 13;1(3):253-8. doi: 10.1016/j.stem.2007.07.019. Epub 2007 Aug 23. PubMed PMID: 18371359.
33: Chan EM, Yates F, Boyer LF, Schlaeger TM, Daley GQ. Enhanced plating efficiency of trypsin-adapted human embryonic stem cells is reversible and independent of trisomy 12/17. Cloning Stem Cells. 2008 Mar;10(1):107-18. doi: 10.1089/clo.2007.0064. PubMed PMID: 18241122.
34: Lensch MW, Daheron L, Schlaeger TM. Pluripotent stem cells and their niches. Stem Cell Rev. 2006;2(3):185-201. Review. PubMed PMID: 17625255.
35: Schlaeger TM, Lensch MW, Taylor PL. Science aside: the trajectory of embryonic stem cell research in the USA. Drug Discov Today. 2007 Apr;12(7-8):269-71. Epub 2007 Mar 1. PubMed PMID: 17395086.
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