2019 / March No. 49 Science, Products, and Information NEXT Interview ips P. 04 StemPro HSC Expansion Medium Prototype P. 05 Heat Stable Recombinant H

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1 19 / March No. 49 Science, Products, and Information NEXT Interview ips P. 4 StemPro HSC Expansion Medium Prototype P. 5 Heat Stable Recombinant Human bfgf P. 6 EVOS M7 Cell Imaging System P. 1 AmpliSeq Transcriptome Gene Expression Assay P xl Latest Version BigDye Terminator v3.1 & v1.1 Cycle Sequencing Kit BigDye XTerminator Purification Kit P. 13 TaqPath 1-Step Multiplex Master Mix P. 14 NanoDrop One

2 ipsinterview 2 3 ips ips ES ES 17 ES ips 8- ips ES ips ips ips ips 1 ips 14 NASH ips ips

3 4 5 StemPro HSC Expansion Medium Prototype CD34 + Heat Stable Recombinant Human bfgf 37bFGF Gibco StemPro HSC Expansion Medium Prototype HSC CD34 + CD9 + CD45RA CD34 + mpb HSC CD34 + CD34 + CD9 + CD45RA CD34 + CFU HSC NEW! bfgf 8 Gibco Heat Stable Recombinant Human bfgf HS bfgf 8% bfgf bfgf >9% FGF NEW! mpbcd34 + HS bfgf 37bFGF A CD34+: Day 7/day (fold) D CD34 + expansion StemPro HSC % CD34 + cells Donor 1 Donor 2 Donor 3 Supplier 1 Supplier 2 Supplier 3 B TNC Day 7/day (fold) TNC expansion StemPro HSC Donor 1 Donor 2 Donor 3 Supplier 1 Supplier 2 Supplier 3 E C Viability (%) at Day 7 Viability of expanded TNCs % CD34 + CD9 + CD45RA cells StemPro HSC Donor 1 Donor 2 Donor 3 Supplier 1 Supplier 2 Supplier 3 Net fluorescence, relative to control (RFU) 3, HS bfgf, 37 C HS bfgf, 4 C 2, 1, 1, , [bfgf] (pg/ml) Net fluorescence, relative to control (RFU) 3, 2, 1, Native bfgf, 37 C Native bfgf, 4 C 1, , [bfgf] (pg/ml) CD34+ at Day 7 (% of TNC count) Donor 1 Donor 2 Donor 3 Donor 1 Donor 2 Donor 3 StemPro HSC Supplier 1 Supplier 2 Supplier 3 1. StemPro HSC Expansion Medium Prototype mpbcd34 + 1% CD34 + mpb HSCStemPro HSC Expansion Medium PrototypemPBCD34 + A TNCB >8% TNC CTNCCD34 + >6% DCD34 + CD9 + CD45RA E StemPro HSC Expansion Medium Prototype3 CD34 + mpb StemPro HSC Expansion Medium Prototype StemPro HSC Basal Medium and 5X Supplement 3 SCF Flt3L TPO IL-3 IL-6 7TNC% Invitrogen Countess II A C D E StemPro HSC Expansion Medium Prototype 5 ml CB183L1 52, CD34 + CD9 + CD45RA at Day 7 (% of TNC count) StemPro HSC Supplier 1 Supplier 2 Supplier 3 1 HS bfgf bfgf PrestoBlue Balb/3T3 1ng/mL HS bfgf bfgf Balb/3T % 2 HS bfgf 72 PrestoBlue Balb/3T34% 1ng/mL HS bfgf bfgf Balb/3T Heat Stable Recombinant Human bfgf 5 ug PHG367 15, bfgf activity, relative to controls, after 72 hrs of heat stress (%) 1 1 Heat Stable bfgf 5 ug PHG368 81,5 1 ug PHG369 19,5 8 6 Native bfgf

4 6 7 EVOS M7 Cell Imaging System Technical Review 3 Invitrogen EVOS M7 Cell Imaging SystemZ- CMOSCMOS D 3 3D 3D 3D 2D 2D3D 2D 3D Invitrogen Celleste Image Analysis Software Invitrogen EVOS Onstage Incubator NEW! LED D 2D Invitrogen Zip Alexa Fluor Antibody Labeling Kits 3D 1 1 HeLa Alexa Fluor 647 Ki67 Zip Alexa Fluor 647 rapid labeling kit Thermo Scientific CellInsight CX7 LZR highcontent analysis HCA Control 2.5µM niclosamide 3T3 6X DAPI GFP RFP 1X GFP RFP 2D live/dead Assay 2 3D 3 2 A549 Invitrogen LIVE/DEAD Cell Imaging Kit CellInsight CX7 LZR HCA 3D HeLa spheroids A549 spheroids 3 24 Invitrogen MitoTracker Orange dye Invitrogen CellEvent Caspase-3/7 Green Detection ReagentCellInsight CX7 LZR HCA FFPEX Ordering information XY EVOS M7 Cell Imaging System 4X Ph GFP & 7,7, 1X FI RFP X FI DAPI X FI 3D A B 4 Invitrogen CellROX Deep Red Reagent HeLa 1 µm A B

5 8 9 PCR PCR NGS K 1K 25K 5K PCR TaqMan TaqMan TaqMan OpenArray 3,72 Ion AmpliSeq Ion AmpliSeq Clariom S Ion Total RNA-Seq Clariom D , >, >, >9, >5, /768 Fast Ion 55 chip Applied Biosystems TaqMan RefSeq 28 PCR Applied Biosystems Clariom 1 pg RNA 1 Clariom D Clariom S Ion Torrent NGS Ion AmpliSeq 1 ng RNA Ion GeneStudio S5 RNA 2 Ion AmpliSeq Transcriptome Assay Plug-in RNA Invitrogen TRIzol Invitrogen PureLink Applied Biosystems MagMAX Invitrogen Cells-to-CT RNA RNADNA mg µg 73 µg 1 8 mg 25 mg.2 ml mg µg RNA MORE INFO Invitrogen SuperScript RT5, SuperScript IV RT cdna RNA Invitrogen Qubit 4 RNA ssdna dsdna Qubit

6 1 11 User's Voice Axcelead Drug Discovery Partners Ion AmpliSeq Transcriptome Assay 1 RNA TaqMan 15Ion AmpliSeq Axcelead Transcriptome Assay Drug Discovery Partners 1 RNAseq Ion Proton 216 Plug-in TaqMan Ion AmpliSeq Transcriptome Assay 2 1ng RNA Ion AmpliSeq Transcriptome Assay, 1ngRNArRNA DNA 96 Axcelead Drug Discovery Partners spin-out 373xl Latest Version Applied Biosystems 373xl ThermoFisher Connect Windows1 373xl LV 373xlLV-1 373xl LV A41978LVUPG-1 BigDye Terminator v3.1 & v1.1 Cycle Sequencing Kit Applied Biosystems BigDye Terminator v3.1 CycleSequencing Kit de novo PCR BAC Applied Biosystems BigDye Terminator v1.1 Cycle Sequencing Kit PCR BigDye Terminator v3.1 Cycle Sequencing Kit , BigDye Terminator v1.1 Cycle Sequencing Kit , NEW! 241, Ion AmpliSeq Transcriptome Gene Expression Assay BigDye XTerminator Purification Kit Ion AmpliSeq Transcriptome Gene Expression Assay, RefSeq 1.1 ngrna RNA Ion GeneStudio S5 Applied Biosystems BigDye XTerminator Purification Kit DNA 1 Ion AmpliSeq Transcriptome Human Gene Expression Kit 24 A , Ion AmpliSeq Transcriptome Mouse Gene Expression Kit 24 A , BigDye XTerminator Purification Kit , 1,

7 12 13 Technical Review User's Voice PowerUP SYBR Green Master Mix PCR 8DNA RNA2 SYBR GreenDNA Ct Applied Biosystems PowerUp SYBR Green Master Mix2Hot Start Dual-Lock Taq DNA Polymerase PowerUp SYBR Green Master MixSYBR Green Master Mix 1 28DNANuclease Free Water No Template Control96 n=3 PowerUp SYBR Green Master Mix1 Ct35 PowerUp SYBR Green Master Mix 1 No Template Control 45Ct 2 VEGFC VEGFC NM_ NM_ A PowerUp SYBR A B 2 2 VEGFC TCF B Ct 3-35 Ct 35- Ct No Template ControlcDNA 5ng61 PCR 2PCR 2 No Template ControlPowerUp SYBR Green Master Mix PowerUp SYBR Green Master Mix PowerUp SYBR A B 21 PoweUp SYBR; 2 A 3 BPCR QuantStudio5 8nM PowerUp SYBR3nM E-Gel 2% agarose gel PowerUp SYBR Green Master Mix 5mL A ,8 web %OFF bp 1 bp pg cdna pg cdna No Template Control No Template Control PCR qpcr 8 2 DNA RNA TaqMan qpcr RT qpcr BRDC 8 2DNA 6RNA 1 ROX TaqPath ROX TaqMan JUN- QSYJUN-QSY BRDC conventional RT-PCR 1 TaqPath 1-Step Multiplex Master MixRNA 1-Step-qPCR MUSTANG PURPLE 4TaqMan ROX 111 BRDC FAM JOE A TAMRA JUN MUSTANG PURPLE FAM RS VIC B 3 TAMRA D JUN MUSTANG PURPLE Passive Referense TaqPath TaqPath 1-Step Multiplex Master Mix 4 PCR D 1 1 qpcr Applied Biosystems TaqPath 1-Step Multiplex Master Mix14RNADNA PCR Applied Biosystems MUSTANG PURPLE TaqPath 1-Step Multiplex Master Mix 5 1 ml A ,8

8 14 15 Technical Review NanoDrop One Thermo Scientific NanoDrop 1-2µL a/b Fe nm 431nm Fe 3+ 6nm NanoDrop 8mg/mL.125mg/mL6 5 6nm A % 1 1 One/One C nm 414nm 524,28M -1 cm kDa 431nm 552,16M -1 cm kDa A6 n=5 n=5 %CV NanoDrop One/One C 75nm 36-5nm MP Biomedicals,LLC J NanoDrop One/One C a/b NanoDrop One 6nm 541nm 576nm NanoDrop One Microvolume UV-Vis Spectrophotometer with Wi-Fi 1 ND-ONE-W 1,75, NanoDrop One C Microvolume UV-Vis Spectrophotometer with Wi-Fi 1 ND-ONEC-W 2,5,?? Title Ph.D Number 31 DNA 86 DNA Ph.D DNA

9 ! INFORMATION Invitrogen Molecular Probes! WEB""!? 2 3D! 3D 3D web 3D NEXT 3? 1! ips 5 11! photographs ryoichi morikawa p2-3 art direction & design: opportune design inc. science writing: momoyo matsuyama / editing: yuko hashimoto NEXT "" For Research Use Only. Not for use in diagnostic procedures. 19 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. TaqMan is a registered trademark of Roche Molecular Systems Inc., used under permission and license. Printed in Japan. LSG52- A193HS TEL FAX

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