10. In the case of iPSC to MK differentiation in bioreactors, or
differentiation from other cell types, please refer to
corresponding MK harvesting steps of the particular protocol.
11. Dilute MK cell suspension in DPBS if the cells are too concentrated, but then take into consideration the dilution factor
in calculations.
References
1. Nakamura S, Takayama N, Hirata S, Seo H,
Endo H, Ochi K, Fujita K, Koike T,
Harimoto K, Dohda T, Watanabe A, Okita K,
Takahashi N, Sawaguchi A, Yamanaka S,
Nakauchi H, Nishimura S, Eto K (2014)
Expandable megakaryocyte cell lines enable
clinically applicable generation of platelets
from human induced pluripotent stem cells.
Cell Stem Cell 14:535–548
2. Sim X, Poncz M, Gadue P, French DL (2016)
Understanding platelet generation from megakaryocytes: implications for in vitro-derived
platelets. Blood 127:1227–1233
3. Cho J (2015) A paradigm shift in platelet transfusion therapy. Blood 125:3523–3525
4. Wang B, Zheng J (2016) Platelet generation
in vivo and in vitro. Springerplus 5:787
5. Moreau T, Evans AL, Vasquez L, Tijssen MR,
Yan Y, Trotter MW, Howard D, Colzani M,
Arumugam M, Wu WH, Dalby A, Lampela R,
Bouet G, Hobbs CM, Pask DC, Payne H,
Ponomaryov T, Brill A, Soranzo N, Ouwehand
WH, Pedersen RA, Ghevaert C (2016) Largescale production of megakaryocytes from
human pluripotent stem cells by chemically
defined forward programming. Nat Commun
7:11208
6. Figueiredo C, Blaszczyk R (2014) Genetically
engineered blood pharming: generation of
HLA-universal platelets derived from CD34+
progenitor cells. J Stem Cells 9:149–161
7. Takayama N, Nishikii H, Usui J, Tsukui H,
Sawaguchi A, Hiroyama T, Eto K, Nakauchi
H (2008) Generation of functional platelets
from human embryonic stem cells in vitro via
ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors. Blood
111:5298–5306
8. Shabrani NC, Khan NF, Kale VP, Limaye LS
(2012) Polyunsaturated fatty acids confer
cryoresistance on megakaryocytes generated
from cord blood and also enhance megakaryocyte production from cryopreserved cord
blood cells. Cytotherapy 14:366–380
9. Ivetic N, Nazi I, Karim N, Clare R, Smith JW,
Moore JC, Hope KJ, Kelton JG, Arnold DM
(2016) Producing megakaryocytes from a
human peripheral blood source. Transfusion
56:1066–1074
10. Borger AK, Eicke D, Wolf C, Gras C,
Aufderbeck S, Schulze K, Engels L,
Eiz-Vesper B, Schambach A, Guzman CA,
Lachmann N, Moritz T, Martin U,
Blasczyk R, Figueiredo C (2016) Generation
of HLA-universal iPSCs-derived megakaryocytes and platelets for survival under refractoriness conditions. Mol Med 22:274–285
11. Wang Y, Hayes V, Jarocha D, Sim X, Harper
DC, Fuentes R, Sullivan SK, Gadue P, Chou
ST, Torok-Storb BJ, Marks MS, French DL,
Poncz M (2015) Comparative analysis of
human ex vivo-generated platelets vs
megakaryocyte-generated platelets in mice: a
cautionary tale. Blood 125:3627–3636
12. Fuentes R, Wang Y, Hirsch J, Wang C,
Rauova L, Worthen GS, Kowalska MA, Poncz
M (2010) Infusion of mature megakaryocytes
into mice yields functional platelets. J Clin
Invest 120:3917–3922
13. Xi J, Zhu H, Liu D, Nan X, Zheng W, Liu K,
Shi W, Chen L, Lv Y, Yan F, Li Y, Xie X,
Wang Y, Yue W, Xu X, Wei X, Zhu J,
Huang X, Pei X (2013) Infusion of megakaryocytic progenitor products generated from
cord blood hematopoietic stem/progenitor
cells: results of the phase 1 study. PLoS One
8:e54941
14. Eicke D, Baigger A, Schulze K, Latham SL,
Halloin C, Zweigerdt R, Guzman CA,
Blasczyk R, Figueiredo C (2018) Large-scale
production of megakaryocytes in microcarriersupported stirred suspension bioreactors. Sci
Rep 8:10146
15. Fuller BJ, Lane N, Benson EE (eds) (2004)
Life in the frozen state. CRC Press, Boca
Raton, FL
16. Mazur P (1988) Stopping biological time. The
freezing of living cells. Ann N Y Acad Sci
541:514–531
17. Mazur P (2010) A biologist’s view of the relevance of thermodynamics and physical chemistry to cryobiology. Cryobiology 60:4–10
Cryopreservation of iPSC-Derived Megakaryocytes
553
differentiation from other cell types, please refer to
corresponding MK harvesting steps of the particular protocol.
11. Dilute MK cell suspension in DPBS if the cells are too concentrated, but then take into consideration the dilution factor
in calculations.
References
1. Nakamura S, Takayama N, Hirata S, Seo H,
Endo H, Ochi K, Fujita K, Koike T,
Harimoto K, Dohda T, Watanabe A, Okita K,
Takahashi N, Sawaguchi A, Yamanaka S,
Nakauchi H, Nishimura S, Eto K (2014)
Expandable megakaryocyte cell lines enable
clinically applicable generation of platelets
from human induced pluripotent stem cells.
Cell Stem Cell 14:535–548
2. Sim X, Poncz M, Gadue P, French DL (2016)
Understanding platelet generation from megakaryocytes: implications for in vitro-derived
platelets. Blood 127:1227–1233
3. Cho J (2015) A paradigm shift in platelet transfusion therapy. Blood 125:3523–3525
4. Wang B, Zheng J (2016) Platelet generation
in vivo and in vitro. Springerplus 5:787
5. Moreau T, Evans AL, Vasquez L, Tijssen MR,
Yan Y, Trotter MW, Howard D, Colzani M,
Arumugam M, Wu WH, Dalby A, Lampela R,
Bouet G, Hobbs CM, Pask DC, Payne H,
Ponomaryov T, Brill A, Soranzo N, Ouwehand
WH, Pedersen RA, Ghevaert C (2016) Largescale production of megakaryocytes from
human pluripotent stem cells by chemically
defined forward programming. Nat Commun
7:11208
6. Figueiredo C, Blaszczyk R (2014) Genetically
engineered blood pharming: generation of
HLA-universal platelets derived from CD34+
progenitor cells. J Stem Cells 9:149–161
7. Takayama N, Nishikii H, Usui J, Tsukui H,
Sawaguchi A, Hiroyama T, Eto K, Nakauchi
H (2008) Generation of functional platelets
from human embryonic stem cells in vitro via
ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors. Blood
111:5298–5306
8. Shabrani NC, Khan NF, Kale VP, Limaye LS
(2012) Polyunsaturated fatty acids confer
cryoresistance on megakaryocytes generated
from cord blood and also enhance megakaryocyte production from cryopreserved cord
blood cells. Cytotherapy 14:366–380
9. Ivetic N, Nazi I, Karim N, Clare R, Smith JW,
Moore JC, Hope KJ, Kelton JG, Arnold DM
(2016) Producing megakaryocytes from a
human peripheral blood source. Transfusion
56:1066–1074
10. Borger AK, Eicke D, Wolf C, Gras C,
Aufderbeck S, Schulze K, Engels L,
Eiz-Vesper B, Schambach A, Guzman CA,
Lachmann N, Moritz T, Martin U,
Blasczyk R, Figueiredo C (2016) Generation
of HLA-universal iPSCs-derived megakaryocytes and platelets for survival under refractoriness conditions. Mol Med 22:274–285
11. Wang Y, Hayes V, Jarocha D, Sim X, Harper
DC, Fuentes R, Sullivan SK, Gadue P, Chou
ST, Torok-Storb BJ, Marks MS, French DL,
Poncz M (2015) Comparative analysis of
human ex vivo-generated platelets vs
megakaryocyte-generated platelets in mice: a
cautionary tale. Blood 125:3627–3636
12. Fuentes R, Wang Y, Hirsch J, Wang C,
Rauova L, Worthen GS, Kowalska MA, Poncz
M (2010) Infusion of mature megakaryocytes
into mice yields functional platelets. J Clin
Invest 120:3917–3922
13. Xi J, Zhu H, Liu D, Nan X, Zheng W, Liu K,
Shi W, Chen L, Lv Y, Yan F, Li Y, Xie X,
Wang Y, Yue W, Xu X, Wei X, Zhu J,
Huang X, Pei X (2013) Infusion of megakaryocytic progenitor products generated from
cord blood hematopoietic stem/progenitor
cells: results of the phase 1 study. PLoS One
8:e54941
14. Eicke D, Baigger A, Schulze K, Latham SL,
Halloin C, Zweigerdt R, Guzman CA,
Blasczyk R, Figueiredo C (2018) Large-scale
production of megakaryocytes in microcarriersupported stirred suspension bioreactors. Sci
Rep 8:10146
15. Fuller BJ, Lane N, Benson EE (eds) (2004)
Life in the frozen state. CRC Press, Boca
Raton, FL
16. Mazur P (1988) Stopping biological time. The
freezing of living cells. Ann N Y Acad Sci
541:514–531
17. Mazur P (2010) A biologist’s view of the relevance of thermodynamics and physical chemistry to cryobiology. Cryobiology 60:4–10
Cryopreservation of iPSC-Derived Megakaryocytes
553