Maintenance and cryopreservation of pluripotent stem cells

Xeno- and serum-free culture of human pluripotent stem cells

  • Reproducible, feeder-free culture up to MACS® GMP grade
  • Flexible feeding schedules that allow for cell culture-free weekends
  • Cryopreservation with high recovery and viability

Application protocols

Discover our different pluripotent stem cell workflows and find the one that fits your experimental needs.

Application data by workflow step

Pluripotent stem cell culture

High-quality human pluripotent stem cell (PSC) lines are the foundation of any successful and reproducible PSC research. Choosing the right cell culture conditions is critical when expanding, passaging, or cryopreserving PSCs. Therefore, we have developed the reliable xeno- and serum-free StemMACS Culture System that is available up to MACS GMP-grade.

Morphology of pluripotent stem cell lines cultured in StemMACS iPS-Brew XF
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StemMACS™ iPS-Brew XF Medium

StemMACS iPS-Brew XF is a xeno-free media
formulation for maintenance and expansion
of highly pluripotent stem cells. Expect typical
pluripotent colony morphology and surface
phenotype, unbiased differentiation potential,
and rapid culture initiation after cryopreservation
with every use.



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Application note
StemMACS iPS-Brew XF enables cell culture-free weekends

In this application note we show that StemMACS iPS-Brew XF enables more flexible feeding schedules, including the possibility of skipping one or even two feeding days, while reliably maintaining PSC cultures. Regardless of the feeding schedule, PSCs retain a characteristic colony morphology, high expansion rates, and high expression levels of pluripotency markers throughout five consecutive passages. 

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Video
Passaging of pluripotent stem cells

See which method we recommend for passaging pluripotent stem cells cultured in StemMACS iPS-Brew XF.


Cryopreservation of PSCs

Optimal storage and cryopreservation is an essential part of many cell culture workflows. StemMACS™ Cryo-Brew is a cryopreservation medium that has been designed for use with xeno- and serum- free culture systems

StemMACS Cryo‑Brew ensures high recovery and viability after cryopreservation of induced PSCs
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StemMACS Cryo-Brew
StemMACS Cryo-Brew ensures high recovery,  viability, and genetic stability after cryopreservation of PSCs.

Phenotypic characterization of PSCs

Thorough characterization of PSCs ensures that your cell lines possess their full differentiation potential. Flow cytometry is a powerful tool to achieve this goal. It provides you with quantitative information about the expression of intracellular as well as surface markers, lets you analyze multiple cell lines at a time, and assesses the efficiency and quality of your differentiation in just a few steps. Take advantage of Miltenyi Biotec’s solutions for flow cytometric analysis and combine them with your favorite cell culture protocol to simplify your characterization workflow and take advantage of the following:

  • Step-by-step protocols for quantitative flow cytometric analysis
  • Recombinantly engineered REAfinity Antibodies for highly reproducible results
  • A simple 7-day assay for functional characterization of pluripotency
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Application note
Multicolor flow cytometric analysis of human pluripotent stem cell cultures

This application note describes  a multicolor flow cytometry protocol that allows for the simultaneous quantification of intracellular and surfacemarkers for easy monitoring of the pluripotency status of human PSC cultures.


Quantitative flow cytometric analysis of trilineage differentiation using the  StemMACS Trilineage Differentiation Kit, human
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StemMACS Trilineage Differentiation Kit, human

Four different iPSC lines were differentiated into ectoderm, mesoderm, and endoderm using the StemMACS Trilineage Differentiation Kit. On day 7, the differentiation cultures were analyzed by flow cytometry. While all lines showed pluripotent differentiation potential, the assay also revealed differences in their propensity to differentiate into certain lineages, e.g. CXCR4+Sox17+ definitive endoderm cells.

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  • Siglec-F
  • Siglec-H
  • SIT1
  • Slan (M-DC8)
  • SLP-76
  • Slug
  • Sox1
  • Sox2
  • Sox17
  • Src pY418
  • SSEA-1
  • SSEA-4
  • SSEA-5
  • SSRP1
  • STAT1 N-terminus
  • STAT1 pS727
  • STAT1 pY701
  • STAT3 pS727
  • STAT4 pY693
  • STAT5a
  • STAT6
  • STAT6 pY641
  • Survivin
  • SUSD2
  • Syk
  • Syk pY348
  • Syndecan-4
  • T-bet
  • TCL1
  • TCR Vα2
  • TCR Vα3.2 (b, c)
  • TCR Vα7.2
  • TCR Vα11.1, 11.2
  • TCR Vα24
  • TCR Vβ1
  • TCR Vβ2
  • TCR Vβ3
  • TCR Vβ4
  • TCR Vβ5
  • TCR Vβ5.1
  • TCR Vβ5.3
  • TCR Vβ6
  • TCR Vβ7.1
  • TCR Vβ7.2
  • TCR Vβ8
  • TCR Vβ8.1, 8.2
  • TCR Vβ9
  • TCR Vβ10
  • TCR Vβ11
  • TCR Vβ12
  • TCR Vβ13.1
  • TCR Vβ13.6
  • TCR Vβ14
  • TCR Vβ16
  • TCR Vβ17
  • TCR Vβ17a
  • TCR Vβ21.3
  • TCR Vβ23
  • TCR Vγ9
  • TCR Vδ1
  • TCR Vδ2
  • TCR Vδ4
  • TCRα/β
  • TCRβ
  • TCRγ/δ
  • TdT
  • Ter-119
  • TGF-β RII
  • THEMIS
  • TIGIT
  • TIM-1
  • TIM-3
  • TIM-4
  • TIMP-3
  • TLT-2
  • TM4SF1
  • TNF-α
  • TNFRH3
  • TOM22
  • TOX
  • TRA-1-60
  • TRA-1-81
  • TRA-1-85 (CD147)
  • TrkA
  • TROP2
  • TSPAN8
  • Under-phospho Rb
  • VEGFR-1
  • Vimentin
  • WIP pS488
  • XCR1
  • ZAP70
  • ZAP70 pY292
  • α-Actinin
  • β-Catenin
  • 1A5
  • 1A8
  • 1B1
  • 1C9-2
  • 1G7.G10
  • 2-2B
  • 2.29
  • 2A2
  • 2A9-1
  • 2C9.G2 (HMβ3-1)
  • 2D10
  • 2E1
  • 2E7
  • 3B2/TA8
  • 3C11
  • 3C12
  • 3G3
  • 3G10
  • 4B4-1
  • 4D11
  • 4E3
  • 4E5
  • 4FR6D3
  • 5-329
  • 5.133
  • 5B1
  • 5C8
  • 5D3
  • 5E8.4
  • 5H10-27
  • 6B11
  • 6D4
  • 6D5
  • 7-239
  • 7A3-3
  • 7C9
  • 7D4
  • 7E.17G9
  • 7E9 
  • 7H11
  • 7H22-E16
  • 8C11
  • 8e11
  • 8F1
  • 8F9
  • 8F10
  • 9E2
  • 10.1.1
  • 10D12
  • 11-26c.2a
  • 11C1
  • 11F2
  • 11F8
  • 11PB6
  • 12G5
  • 13G3
  • 14B11
  • 15E8
  • 16-10A1
  • 17A2
  • 17B5-1H1
  • 18d3
  • 20/70
  • 20d5
  • 22E5
  • 23C6
  • 24D2
  • 24F
  • 25-D1.16
  • 29A1.4.9
  • 30-H12
  • 30F11
  • 32D12
  • 43A4E1
  • 44F9
  • 45-15
  • 53-6.7
  • 53-7.3
  • 67A4
  • 67D2
  • 87G
  • 90.4
  • 93
  • 97C5
  • 104-2
  • 123R3
  • 145-2C11
  • 145/15
  • 187.1
  • 188B
  • 191B8
  • 293C3
  • 390
  • 429 (MVCAM.A)
  • 541-10B2
  • 551.3D3
  • 555
  • A1
  • A2F10
  • A3C6E2
  • A7R 34
  • A20
  • AC102
  • AC104.3E3
  • AC106
  • AC122
  • AC128
  • AC133
  • AC136
  • AC144
  • AC145
  • ACT35
  • AD2
  • AD5-8E7
  • AD5-14H12
  • AD5-17F6
  • AF12-7H3
  • AF29-4D12
  • AFS98 
  • APA5
  • APB5
  • ATM-2
  • Avas12
  • B3B4
  • B4B6
  • BAT221
  • BB27
  • Ber-ACT 8
  • Bio3-18E7
  • BM16
  • BT3.1
  • BV10A4H2
  • BVD4-1D11
  • BW135/80
  • BW242/412
  • BW264/56
  • C9B7W
  • C363-16A
  • cA2
  • caa7-9G8
  • caa8-28H10
  • CD7-6B7
  • CH3A4A7
  • Cr24.1
  • CRA1
  • CW-1.2
  • CX5
  • Cy34.1
  • CZ8-23G1
  • D7
  • D7715A7
  • DA03-22H2.8
  • DA04-10E8.9
  • DB105
  • DCN47.5
  • DCN228
  • DD-1
  • DF-T1
  • DG3
  • DJR1
  • DJR3
  • DN254
  • DT5D3
  • DX2
  • DX5
  • DX9
  • DX11
  • DX27
  • E11
  • EAT2
  • EP-1
  • ES8-20E6
  • ES22-10D8
  • ES26-5E12.4
  • ES26-14D1.11
  • ES26-15B7.3
  • F38-2E2
  • FA-11
  • FGK45.5
  • FM95
  • FM276
  • FN50
  • FR3-16A11
  • FR10-11G9
  • FR 70 
  • GHI/61.1
  • GHI/75
  • GK1.5
  • GL3
  • GoH3
  • H1.2F3
  • H4B4
  • H5C6
  • H35-17.2
  • H44
  • H100-27.R55
  • H194-112
  • HA2-7B1
  • HB5
  • HB14
  • HB15
  • HBF-719
  • hC3aRZ8
  • HD30
  • HEA-125
  • HECA-452
  • HI149
  • His51
  • HK5.3
  • Hly9.1.25
  • HM48-1
  • HM79-12
  • HMD4-1
  • HMN2-35
  • HMß1-1
  • HMα5-1
  • HSL96
  • HTF-1
  • IB6
  • IgD26
  • IM7.8.1
  • IS5-20C4
  • IS5-21F5
  • IS7-24C7
  • IS11-3B2.2.3
  • IS11-8E10
  • IS11-21E11
  • IS11-24D5
  • ITEM-4
  • JC5-1
  • JD3
  • JES1-39D10
  • JES3-9D7
  • JES5-16E3
  • JF05-1C2.4.1
  • JJC11.6
  • JS11
  • KJ1-26
  • L11
  • LG.3A10
  • LN2-9C4
  • LT2
  • LT19
  • LT20
  • LT27:295
  • M-L23.4
  • M-T271
  • M-T411
  • M-T466
  • M1/69
  • M1/70.15.11.5
  • M3/38
  • M3/84
  • M5/114.15.2
  • M18/2
  • MAR-1
  • MB9-3G8
  • MB10-5C4
  • MB15-18C9
  • MB22-9F5
  • MD5-1-3C2
  • ME-9F1
  • ME20.4-1.H4
  • MEL14-H2.100
  • MFL3
  • MHD1-314
  • MHD4-46
  • MHR73-11
  • MIH12
  • MIH18
  • MIH37
  • MJ4-27G12
  • MJ7/18
  • MRα4-1
  • MWReg30
  • MZ3
  • MZ18-23C8
  • MZ18-24A9
  • N7.48 A
  • N418
  • NKVFS1
  • NLDC-145
  • NOK-1
  • OX-104
  • PD1.3.1.3 
  • PK136
  • PO3.3
  • PV404.19
  • R1-2
  • R1.302
  • RA3-6B2
  • RB6-8C5
  • REA100
  • REA101
  • REA103
  • REA107
  • REA108
  • REA109
  • REA110
  • REA111
  • REA112
  • REA113
  • REA114
  • REA115
  • REA118
  • REA119
  • REA120
  • REA121
  • REA122
  • REA123
  • REA124
  • REA125
  • REA126
  • REA127
  • REA128
  • REA129
  • REA134
  • REA138
  • REA141
  • REA142
  • REA143
  • REA144
  • REA145
  • REA146
  • REA147
  • REA148
  • REA151
  • REA153
  • REA154
  • REA158
  • REA161
  • REA163
  • REA164
  • REA165
  • REA167
  • REA168
  • REA170
  • REA171
  • REA173
  • REA175
  • REA176
  • REA179
  • REA181
  • REA185
  • REA187
  • REA188
  • REA190
  • REA191
  • REA192
  • REA193
  • REA194
  • REA195
  • REA196
  • REA198
  • REA199
  • REA200
  • REA202
  • REA203
  • REA204
  • REA205
  • REA206
  • REA207
  • REA208
  • REA209
  • REA211
  • REA213
  • REA214
  • REA215
  • REA217
  • REA218
  • REA219
  • REA220
  • REA221
  • REA222
  • REA223
  • REA226
  • REA227
  • REA228
  • REA230
  • REA231
  • REA232
  • REA233
  • REA234
  • REA236
  • REA237
  • REA238
  • REA241
  • REA244
  • REA245
  • REA247
  • REA250
  • REA252
  • REA254
  • REA256
  • REA257
  • REA258
  • REA259
  • REA260
  • REA261
  • REA263
  • REA264
  • REA265
  • REA266
  • REA269
  • REA273
  • REA274
  • REA276
  • REA277
  • REA279
  • REA280
  • REA282
  • REA284
  • REA286
  • REA287
  • REA289
  • REA290
  • REA292
  • REA293
  • REA296
  • REA298
  • REA299
  • REA300
  • REA301
  • REA302
  • REA303
  • REA304
  • REA306
  • REA308
  • REA309
  • REA311
  • REA312
  • REA313
  • REA315
  • REA316
  • REA318
  • REA319
  • REA321
  • REA322
  • REA325
  • REA327
  • REA329
  • REA332
  • REA336
  • REA337
  • REA339
  • REA340
  • REA343
  • REA344
  • REA347
  • REA349
  • REA351
  • REA354
  • REA359
  • REA360
  • REA361
  • REA363
  • REA364
  • REA366
  • REA368
  • REA372
  • REA374
  • REA377
  • REA378
  • REA381
  • REA382
  • REA384
  • REA386
  • REA387
  • REA388
  • REA389
  • REA391
  • REA392
  • REA393
  • REA395
  • REA396
  • REA398
  • REA400
  • REA402
  • REA405
  • REA408
  • REA409
  • REA410
  • REA412
  • REA413
  • REA414
  • REA419
  • REA420
  • REA421
  • REA422
  • REA423
  • REA425
  • REA426
  • REA428
  • REA429
  • REA431
  • REA437
  • REA440
  • REA441
  • REA442
  • REA443
  • REA444
  • REA445
  • REA446
  • REA447
  • REA448
  • REA450
  • REA452
  • REA453
  • REA456
  • REA459
  • REA460
  • REA461
  • REA464
  • REA465
  • REA466
  • REA467
  • REA469
  • REA470
  • REA471
  • REA472
  • REA476
  • REA478
  • REA479
  • REA482
  • REA484
  • REA489
  • REA490
  • REA492
  • REA493
  • REA494
  • REA495
  • REA496
  • REA497
  • REA499
  • REA501
  • REA502
  • REA503
  • REA504
  • REA505
  • REA508
  • REA509
  • REA510
  • REA512
  • REA513
  • REA517
  • REA518
  • REA519
  • REA523
  • REA524
  • REA526
  • REA527
  • REA528
  • REA531
  • REA532
  • REA535
  • REA538
  • REA540
  • REA541
  • REA542
  • REA543
  • REA545
  • REA546
  • REA547
  • REA550
  • REA553
  • REA554
  • REA557
  • REA558
  • REA559
  • REA560
  • REA561
  • REA562
  • REA563
  • REA564
  • REA566
  • REA568
  • REA570
  • REA571
  • REA572
  • REA573
  • REA574
  • REA577
  • REA578
  • REA581
  • REA585
  • REA587
  • REA589
  • REA591
  • REA592
  • REA594
  • REA596
  • REA598
  • REA599
  • REA600
  • REA601
  • REA602
  • REA603
  • REA604
  • REA605
  • REA606
  • REA607
  • REA608
  • REA610
  • REA611
  • REA612
  • REA613
  • REA614
  • REA615
  • REA616
  • REA617
  • REA618
  • REA619
  • REA620
  • REA621
  • REA623
  • REA624
  • REA625
  • REA627
  • REA629
  • REA630
  • REA631
  • REA633
  • REA634
  • REA635
  • REA637
  • REA638
  • REA639
  • REA640
  • REA641
  • REA642
  • REA645
  • REA646
  • REA648
  • REA649
  • REA651
  • REA652
  • REA654
  • REA657
  • REA661
  • REA662
  • REA663
  • REA664
  • REA665
  • REA669
  • REA670
  • REA671
  • REA672
  • REA674
  • REA675
  • REA677
  • REA679
  • REA680
  • REA683
  • REA684
  • REA686
  • REA687
  • REA688
  • REA689
  • REA690
  • REA692
  • REA693
  • REA694
  • REA695
  • REA698
  • REA699
  • REA701
  • REA706
  • REA707
  • REA709
  • REA711
  • REA713
  • REA714
  • REA715
  • REA717
  • REA718
  • REA722
  • REA723
  • REA724
  • REA727
  • REA729
  • REA730
  • REA733
  • REA734
  • REA736
  • REA737
  • REA738
  • REA739
  • REA740
  • REA741
  • REA743
  • REA744
  • REA745
  • REA747
  • REA748
  • REA749
  • REA750
  • REA752
  • REA753
  • REA754
  • REA755
  • REA757
  • REA758
  • REA759
  • REA760
  • REA761
  • REA762
  • REA764
  • REA765
  • REA768
  • REA769
  • REA770
  • REA771
  • REA772
  • REA773
  • REA774
  • REA775
  • REA776
  • REA777
  • REA778
  • REA779
  • REA780
  • REA781
  • REA782
  • REA784
  • REA785
  • REA786
  • REA787
  • REA788
  • REA789
  • REA790
  • REA791
  • REA792
  • REA793
  • REA794
  • REA795
  • REA796
  • REA797
  • REA798
  • REA800
  • REA801
  • REA802
  • REA803
  • REA804
  • REA805
  • REA806
  • REA807
  • REA808
  • REA809
  • REA810
  • REA811
  • REA812
  • REA813
  • REA815
  • REA816
  • REA817
  • REA818
  • REA819
  • REA820
  • REA822
  • REA823
  • REA824
  • REA826
  • REA827
  • REA828
  • REA829
  • REA831
  • REA832
  • REA833
  • REA834
  • REA835
  • REA836
  • REA838
  • REA839
  • REA840
  • REA841
  • REA842
  • REA843
  • REA844
  • REA845
  • REA846
  • REA847
  • REA848
  • REA849
  • REA850
  • REA851
  • REA852
  • REA853
  • REA855
  • REA856
  • REA857
  • REA858
  • REA859
  • REA860
  • REA861
  • REA862
  • REA863
  • REA864
  • REA865
  • REA867
  • REA868
  • REA869
  • REA870
  • REA871
  • REA872
  • REA873
  • REA874
  • REA875
  • REA876
  • REA877
  • REA878
  • REA879
  • REA880
  • REA881
  • REA882
  • REA884
  • REA886
  • REA887
  • REA889
  • REA890
  • REA892
  • REA893
  • REA894
  • REA895
  • REA896
  • REA897
  • REA898
  • REA899
  • REA901
  • REA902
  • REA903
  • REA905
  • REA906
  • REA907
  • REA909
  • REA910
  • REA911
  • REA912
  • REA915
  • REA916
  • REA918
  • REA923
  • REA932
  • REA934
  • REA936
  • REA937
  • REA940
  • REA941
  • REA942
  • REA943
  • REA945
  • REA946
  • REA947
  • REA948
  • REA950
  • REA951
  • REA953
  • REA954
  • REA955
  • REA956
  • REA957
  • REA958
  • REA959
  • REA960
  • REA961
  • REA962
  • REA963
  • REA964
  • REA965
  • REA967
  • REA968
  • REA969
  • REA970
  • REA971
  • REA972
  • REA975
  • REA976
  • REA977
  • REA978
  • REA979
  • REA980
  • REA981
  • REA983
  • REA984
  • REA985
  • REA988
  • REA989
  • REA990
  • REA991
  • REA994
  • REA995
  • REA996
  • REA997
  • REA998
  • REA999
  • REA1000
  • REA1002
  • REA1003
  • REA1005
  • REA1006
  • REA1007
  • REA1008
  • REA1010
  • REA1012
  • REA1013
  • REA1014
  • REA1015
  • REA1016
  • REA1017
  • REA1018
  • REA1019
  • REA1020
  • REA1021
  • REA1023
  • REA1026
  • REA1028
  • REA1030
  • REA1031
  • REA1034
  • REA1036
  • REA1037
  • REA1039
  • REA1040
  • REA1041
  • REA1042
  • REA1043
  • REA1045
  • REA1046
  • REA1047
  • REA1048
  • REA1049
  • REA1051
  • REA1053
  • REA1054
  • REA1055
  • REA1056
  • REA1057
  • REA1058
  • REA1061
  • REA1062
  • REA1065
  • REA1066
  • REA1067
  • REA1070
  • REA1071
  • REA1075
  • REA1076
  • REA1078
  • REA1080
  • REA1081
  • REA1082
  • REA1083
  • REA1095
  • REA1098
  • REA1100
  • REA1103
  • REA1106
  • RG7/1.30
  • RG7/11.1
  • RG11/39.2
  • RM2-5
  • RM134L
  • RMT1-17
  • RP/14
  • S5/1
  • S43.10
  • SN4 C3-3A2
  • SN13
  • T6D11
  • TB03
  • TC11-18H10
  • Ter-119
  • TET2
  • TKS-1
  • TLR 3.7
  • TM-β1
  • TS1/18
  • TS2/7
  • TS2/9
  • TS2/16
  • TY/11.8
  • TÜK4
  • UCHL1
  • UCHT2 
  • UP-D2
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