<p>Supplementary Figures to</p><p>Variable resistance to freezing and thawing of CD34 positive stem cells and lymphocyte subpopulations in leukapheresis products</p><p>Supplementary Figure 1A. Analysis report of CD34+ cell and CD45+ cell viability measurement. Flow cytometry analysis was performed on a single platform using CD45 </p><p>FITC/CD34 PE antibodies, 7-AAD viability dye, and Trucount beads (BD Biosciences, San Jose, </p><p>CA) according to the modified ISHAGE protocol. A report of a representative post thaw product sample is shown. Data.006 Enter These Values Data (Total .006 CD45) Trucount = 48450.00 No gate</p><p>R2 Dilution Factor = R1 R5</p><p>0 1 2 3 10.00 Sample 0 1 2 3 10 10 10 10 10 10 10 10 4 4 10 10 CD34 PE Volume = CD45 FITC Data.006 Data.006 R2*G1 50.00</p><p>R3 File: Data.006 Sample ID: 15/134-4 Patient ID: XXXXXXXXXX Trucount: 48450 Dilution Factor: 10 Sample Volume: 50 Acquisition Date: 04- Dec-15 Total Events: 159356 R1</p><p>0 1 2 3 10 10 10 10 0 200 400 600 800 4 1000 10 FSC-Height Beads 23682 14.86 CD45 FITC Total CD34 1667 1.05 Data.006 Viable Lymphs 25070 15.73 Data.006 Total CD45 134952 84.69 No gate Debris 0 0.00</p><p>R6 Inspect All Dotplots Results Total CD45 = 55218.52 /uL</p><p>0 200 400 600 800 0 1 2 3 10 10 10 10 1000 CD45 Viability = 53.02 % 4 FSC-Height 10 CD45 FITC Data.006 Viable CD34 = 342.89 /uL Data.006 Viable cells R1*R2*R3 Viable CD45 = 29274.59 /uL</p><p>R8 Total CD34 = 682.09 /uL</p><p>R8</p><p>0 1 2 3 CD34 Viability = 50.27 % 10 10 10 10 0 1 2 3 4 10 10 10 10 10 4 7-AAD 10 Viable CD34 of Viable CD45 = 1,17 % 7-AAD</p><p>Total CD34 of Total CD45 = 1.24 %</p><p>Page 1 Supplementary Figure 1B. Analysis report of lymphocyte subpopulation measurement. Flow cytometry analysis was performed in a dual platform assay using TriTest reagents (CD3/CD4/C45, </p><p>CD3/CD8/CD45, CD3/CD19/CD45, CD3/CD16+C56/CD45; BD Biosciences) and BD MultiSET software for data analysis.</p><p>Reports of a representative post thaw product sample are shown. MultiSET™ Physician Report Director: Prof. Dr. J. Oldenburg Software: MultiSET V1.1.2 Operator: LE Cytometer: FACScan (#81343)</p><p>Sample 6 Date Acquired: Die, 17. Nov Name: 15/134-4 2015 13:47 Uhr Sample XXXXXXXX Date Analyzed: ID: Case XX Number: 3 Color Die, 17. Nov 2015 Panel TBNK Reference Range Name: WBC: Type: BD User 3 Inputs: 6.1 x10 /µL Percent Lymphs: 25.6 Abs Cnt Resu % (cells/µL) Reference lt / Range Nam R 55% e a t 84% i o 690</p><p>2540</p><p>31%</p><p>60%</p><p>410</p><p>1590</p><p>13% 41%</p><p>190 1140</p><p>6% 25%</p><p>90 660</p><p>5% 27%</p><p>90 590</p><p>Lymphosum: 99.9 CD3 % Lymph 3 Difference: CD3 1001 - 1050 Abs Cnt Range: 1.41 T Helper/Suppresso r Ratio: Comments: Laboratory Director: MultiSET™ Lab Report Prof. Dr. J. Oldenburg Director: LE Software: MultiSET V1.1.2 Operator: Cytometer: FACScan (#81343)</p><p>S D Die, 17. Nov 2015 13:47 a at Uhr m e p A l c e q ui N re a d: m e : 6 Sam D Die, 17. Nov 2015 ple a ID: t e</p><p>A n a l y z e d : Case Ref. Range Type: BD Number: XXXXXXX XXXX Panel 3 User 6 Lymphs Name: Inputs: . (%): 25.6 C 1 o WBC l (x1000): o r T B N K</p><p>CD3/ D Data 15/134-4 06.01 CD4/ a File: CD45 t a</p><p>S e t [ 1</p><p>] Reagent Lot ID: 46970 Attr Def File: 3/4/45 Tri v2.0 Events Acquired: 15000</p><p>C a s e</p><p>N u m b e r :</p><p>P D 1 a a 5 n t / e a 1 3 l F 4 il - N e 4 a : 0 m 6 e . : 0 2</p><p>S a m p l e</p><p>I D : CD3/CD19/CD45 Data Set [ 1 ] Data File: 15/134-4 06.03 Re Attr age Def nt File: Lot 3/19/ ID: 45 619 Tri 81 v2.0 Eve nts Acq uire d: 165 00</p><p>C D 15/134-4 D a 06.04 3 t a / C F D il e 1 : 6 + 5 6 / C D 4 5 Re Attr age Def nt File: Lot 3/16+ ID: 56/45 257 Tri 17 v2.1 Eve nts Acq uire d: 165 00 9 3 9 1001 - 1050 . 9 1.4</p><p>Multi-tube QC /Suppressor L Ratio: y m p h o s u m : C D 3 % L y m p h Di ff er e n c e: C D 3 A b s C nt R a n g e: T H el p er after thawing (gray). </p><p>The right peak shows </p><p> the CD3+ events.</p><p>Supplementary Figure</p><p>2. Overlay analysis of </p><p>CD3+ events in cryopreserved apheresis products. </p><p>CD3+ events of apheresis products from two healthy volunteers were compared in samples taken before freezing (black) and Supplementary Figure</p><p>3. NK cell post thaw recovery and liquid storage time. Post thaw</p><p>CD16+CD56+ cell recovery was determined in 90 cryopreserved apheresis products. Liquid storage time shows the time between end of apheresis and end of cryopreservation.</p>