Stem cell research solutions and support

Stem cell research solutions and support

More resources than any other supplier

r& d live cha t online forums field applic ation specialists webinars posters protocols tech tips

Stem cell research can be time-consuming.

Finding support shouldn’t be.

Go to lifetechnologies.com/pscresources to check out protocols,

FAQs, publications, scientific posters, videos, and upcoming events.

That’s why Life Technologies gives you easy access to

70+ proven protocols, 2,500+ peer-reviewed publications,

how-to videos, and webinars from leaders in the stem

cell community. And, when you have a question, technical

support is always available by phone, email, live chat, or

in person. It’s all about getting you everything you need to

ensure stem cell success.

Technical resources

Contents

4

Pluripotent stem cells

4 Culture

8 Tools for reprogramming

10 Detection

11 Analysis

12

Mesenchymal stem cells

12 Culture

14 Differentiation

15 Detection

16

Neural stem cells

16 Culture

16 Differentiation

17

Hematopoietic stem cells

17 Culture

lentivirus—sendai virus—episomal vectors

Pluripotent stem cells (PSCs)

Culture

Pluripotent stem cell (PSC) research requires careful attention

to culture conditions. To find the right solution for the right

environment, tap into peer-reviewed publications, protocols,

instructional videos, and stem cell experts. From KnockOut

_Serum

Replacement to Essential 8

_{Medium and cell therapy research}

media, Gibco

_{products deliver culture with confidence.}

lifetechnologies.com/culturepsc

KnockOut

_{Serum Replacement (KSR)}

Well-referenced, proven standard

Knockout

_{serum replacement and}

basal media deliver confidence, with over

2,400 peer-reviewed publications.

Cat. No. 10828010, 10828028

Data image for KnockOut™ _{SR XenoFree CTS}™ _{Kit provided by}

Dr. Hidenori Akutsu, National Center for Child Health and Development, Tokyo, Japan.

Which pluripotent stem cell culture medium is right for you?

The standard for serum-free, feeder-based culture

of ESCs and iPSCs from multiple species Feeder-free culture with flexibility to optimize bFGF concentration and maintain differentiation capacity of PSCs Optimal feeder-free culture of all human ESC and iPSC lines with minimal lot-to-lot variability Fully documented xeno-free PSC culture reducing the burden in qualifying reagents, facilitating transition, from the bench to the clinic

Product name KnockOut™ _{Serum Replacement StemPro}® _{hESC SFM Essential 8}™ _{Medium KnockOut}™ _CTS™ _{XenoFree ESC/iPSC Kit}

cat. no. 10828010, 10828028 a1000701 a14666sa a1448801, a1448701

Recommended applications Multiple species; feeder culture; reprogramming on

feeders; differentiation of PSCs Human; feeder-free culture Human; feeder-free, xeno-free culture; feeder-free reprogramming Human; feeder-based and feeder-free xeno-free culture for cell therapy research

lot-to-lot variability medium medium low medium

cell morphology compact colonies with smooth edges;

high nuclear to cytoplasmic ratio compact colonies with smooth edges; some colonies will have undefined borders with spikey edges compact colonies with smooth edges; high nuclear to cytoplasmic ratio compact colonies with smooth edges; some colonies will have undefined borders with spikey edges components/unit 1 x 100 ml or 500 ml 1 x 100 ml stempro® _{hesc supplement}

1 x 500 ml dmem/f12 with Glutamax™ _medium

1 x 50 ml 25% bsa

1 x 10 ml essential 8™ _supplement

1 x 500 ml dmem/f-12 (ham) 1:1 1 x 100 ml Knockout

™ _{sr xenofree cts}™

1 x 500 ml Knockout™ _{dmem cts}™

1 x 5 ml Knockout™ _{sr Gf cocktail cts}™

additional components required for

complete medium dmem/f12 (1x) with Glutamax

™ _{medium, neaa,}

bfGf; 2-mercaptoethanol; for a cost-effective option, choose the Knockout™ _{esc/ipsc media Kit}

bfGf and 2-mercaptoethanol no additional components required bfGf cts™ _{and 2-mercaptoethanol}

price/500 ml of complete medium $$ individually

$ for kit $$ $ $$$$

stability of complete medium stable at 4°c for 2 weeks stable at 4°c for 2 weeks stable at 4°c for 2 weeks stable at 4°c for 2 weeks specifically designed for cell therapy

research no no no Yes

available support resources Protocols for culturing PSCs, publications,

hands-on training, product finder Protocols for culturing PSCs Protocols for culturing PSCs, FAQs, hands-on training Protocols for culturing PSCs, FAQs, product documentation

StemPro

_{hESC SFM}

Flexible feeder-free culture

stempro

_{hesc sfm optimizes bfGf}

concentration and maintains the

differentiation capability of pscs.

Cat. No. A1000701

Essential 8

_Medium

Feeder-free culture for all

essential 8

_{medium contains only the}

necessary components for optimal psc

culture for every step of your research.

Cat. No. A14666SA

KnockOut

_{SR XenoFree CTS}

_Kit

Traceability for cell therapy

research

the Knockout

_{sr xenofree cts}

_Kit

reduces the burden in qualifying reagents,

facilitating transition from the bench to

the clinic.

Cat. No. A1448801, A1448701

Which pluripotent stem cell culture medium is right for you?

The standard for serum-free, feeder-based culture

of ESCs and iPSCs from multiple species Feeder-free culture with flexibility to optimize bFGF concentration and maintain differentiation capacity of PSCs Optimal feeder-free culture of all human ESC and iPSC lines with minimal lot-to-lot variability Fully documented xeno-free PSC culture reducing the burden in qualifying reagents, facilitating transition, from the bench to the clinic

Product name KnockOut™ _{Serum Replacement StemPro}® _{hESC SFM Essential 8}™ _{Medium KnockOut}™ _CTS™ _{XenoFree ESC/iPSC Kit}

cat. no. 10828010, 10828028 a1000701 a14666sa a1448801, a1448701

Recommended applications Multiple species; feeder culture; reprogramming on

feeders; differentiation of PSCs Human; feeder-free culture Human; feeder-free, xeno-free culture; feeder-free reprogramming Human; feeder-based and feeder-free xeno-free culture for cell therapy research

lot-to-lot variability medium medium low medium

cell morphology compact colonies with smooth edges;

high nuclear to cytoplasmic ratio compact colonies with smooth edges; some colonies will have undefined borders with spikey edges compact colonies with smooth edges; high nuclear to cytoplasmic ratio compact colonies with smooth edges; some colonies will have undefined borders with spikey edges components/unit 1 x 100 ml or 500 ml 1 x 100 ml stempro® _{hesc supplement}

1 x 500 ml dmem/f12 with Glutamax™ _medium

1 x 50 ml 25% bsa

1 x 10 ml essential 8™ _supplement

1 x 500 ml dmem/f-12 (ham) 1:1 1 x 100 ml Knockout

™ _{sr xenofree cts}™

1 x 500 ml Knockout™ _{dmem cts}™

1 x 5 ml Knockout™ _{sr Gf cocktail cts}™

additional components required for

complete medium dmem/f12 (1x) with Glutamax

™ _{medium, neaa,}

bfGf; 2-mercaptoethanol; for a cost-effective option, choose the Knockout™ _{esc/ipsc media Kit}

bfGf and 2-mercaptoethanol no additional components required bfGf cts™ _{and 2-mercaptoethanol}

price/500 ml of complete medium $$ individually

$ for kit $$ $ $$$$

stability of complete medium stable at 4°c for 2 weeks stable at 4°c for 2 weeks stable at 4°c for 2 weeks stable at 4°c for 2 weeks specifically designed for cell therapy

research no no no Yes

available support resources Protocols for culturing PSCs, publications,

hands-on training, product finder Protocols for culturing PSCs Protocols for culturing PSCs, FAQs, hands-on training Protocols for culturing PSCs, FAQs, product documentation

KnockOut

_{ESC/iPSC Media Kit}

a cost-effective and convenient way to purchase

the reagents required for feeder-based culture of

human and nonhuman embryonic stem cells (escs)

or induced pluripotent stem cells (ipscs). this

kit includes dmem/f12 with Glutamax

_medium,

Knockout

_{serum replacement, and fGf-basic.}

Cat. No. A1412901, A1413001

Pluripotent stem cells (PSCs)

Complementary culture products

Which extracellular matrix is right for you?

we provide extracellular matrices (ecms) that are designed to minimize adaptation time, maximize cell performance, and help meet regulatory requirements. whether you are culturing your cells on feeder layers or require feeder-free, xeno-free conditions for more clinical applications, Gibco® _{products provide you with}

all the choices to culture your pscs in the right environment.

Most commonly used basement membrane matrix for ESC and iPSC cultures

Optimized for use with

Essential 8™ _Medium Optimal for clinical research _{applications that require human}

origin materials only

Product name Geltrex® _{LDEV-Free hESC Qualified Vitronectin (VTN-N) CELLstart}™ _CTS™

cat. no. a1413301 a14701sa a1014201

Recommended applications Feeder and feeder-free culture Feeder-free and xeno-free culture Feeder-free, xeno-free culture

recommended medium pairing stempro® _{hesc sfm essential 8}™ _{medium Knockout}™ _{sr xenofree cts}™

-supplemented medium source origin murine human (recombinant) blend of human native proteins

defined no Yes Yes

storage temperature frozen (–20 to –80°c) frozen (–20 to –80° c) refrigerate (2–8°c) plate preparation time 1 hr incubation at 37°c 1 hr incubation at room temperature 1–2 hrs incubation at 37°c shelf life of prepared plate 2 weeks at 2–8°c 1 week at 2–8°c 1–7 days at 2–8°c (tightly sealed)

unit size 1 ml or 5 ml 1 ml 2 ml

number of plates coated/unit 16 x 6-well plates from 1 ml vial

diluted 1:100 16 x 6-well plates from 1 ml 22 x 6-well plates when diluted 1:50

relative price/6-well plate $ $$ $$$

attachment factor (0.1% Gelatin, cat. no. s006100) may be used for feeder-layer attachment. $—inexpensive $$—moderate $$$—expensive $$$$—exclusive

Which passaging method is right for you?

whether you prefer to passage your cells enzymatically or manually, Gibco® _{media provides you with choices for psc subculture.}

Recommended for clump passaging in feeder and feeder-free cultures

Standard reagent for clump pas-saging in feeder and feeder-free cultures Quick and gentle dissociation, recommended for single-cell passaging Recommended for clump passaging with feeder-free Essential 8™ Medium

Optimal for clinical research applications that require human origin materials only

Quick and easy manual passaging tool for feeder and feeder-free cultures

Product name Dispase Collagenase IV Accutase® _UltraPure™ _0.5M

EDTA TrypLE

™ _{Select CTS}™ _StemPro®

EZPassage™

Disposable Stem Cell Passaging Tool

cat. no. 17105041 17104019 a1110501 15575020 a1285901 23181010

Recommended

applications Passaging in clumps Passaging in clumps Dissociation to single cells Passaging in clumps Passaging in clumps or single-cell passaging

Mechanical passaging in clumps

source origin Bacillus polymyxa Clostridium

histolyticum marine chemical (animal origin–free (aof)) recombinant enzyme (aof) na

form lyophilized powder lyophilized powder liquid, ready to use liquid, dilute 1,000x liquid, ready to use individually wrapped storage temperature 2–8°c 2–8°c 2–8°c room temperature room temperature room temperature shelf life of liquid

format 2 weeks 2 weeks 2 years 2 years 2 years na

inactivation method inhibition by dilution (no inhibitors needed) inhibition by dilution (no inhibitors needed) inhibition by dilution (no inhibitors needed) inhibition by dilution (no inhibitors needed) inhibition by dilution (no inhibitors needed) na

unit size 5 g 1 g 100 ml 4 x 100 ml 100 ml box of 10 units

optimized

concentration for use 2 mg/ml 1 mg/ml for feeder-dependent, 10 mg/ ml for feeder-free

1x solution 0.5 mm 1x solution na number of plates

passaged/unit 1,250 x 60 mm plates 500 x 60 mm plates at 1 mg/ml, 50 x 60 mm plates at 10 mg/ml 50 x 60 mm plates 50,000 x 60 mm plates (100 ml at 1,000x dilution) 50 x 60 mm plates 10 plates relative price/60 mm plate $ $ $$ $ $$ $$$

$—inexpensive $$—moderate $$$—expensive $$$$—exclusive

An iPSC colony cut into evenly sized pieces using the StemPro® _EZPassage™ _Disposable

Tools for reprogramming

Selecting the right reprogramming tool to generate induced pluripotent stem cells (iPSCs) can leave you with questions. And no

one has more answers than Life Technologies. From traditional methods (lentivirus) to nonintegrating technologies (Sendai virus

and episomal vectors), Invitrogen

_{products provide the breadth and depth you depend on. That means more solutions to meet}

your safety, efficiency, and affordability needs.

lifetechnologies.com/cellreprogram

iPSC colonies generated using Episomal iPSC Reprogramming Vectors Express TRA-1-81 (green) at Day 24.

(A-B) 200,000 primary human fibroblasts were reprogrammed with (A) the CytoTune®_-iPS

Sendai Reprogramming Kit and (B) lentivirus. Twenty-eight days post-reprogramming, ~245 colonies were observed from CytoTune® _{reprogramming, while the lentivirus reprogramming}

generated ~10 colonies. (C-D) 50,000 primary human fibroblasts were reprogrammed with (C) the CytoTune®_{-iPS Sendai Reprogramming Kit and (D) mRNA. Twenty-eight days}

post-reprogramming, ~130 iPSC colonies were observed from CytoTune® _{reprogramming, while 11}

iPSC colonies were observed from mRNA reprogramming.

Pluripotent stem cells (PSCs)

A. CytoTune®_{-iPS Sendai Reprograming Kit B. Lentivirus}

C. CytoTune®_{-iPS Sendai Reprogramming Kit D. mRNA}

Lentiviral iPSC reprogramming

particles

Well-referenced, flexible

technology

choose from eight different high-titer

lentiviral ipsc reprogramming particles

for your custom ipsc experiments.

lifetechnologies.com/lenti

Episomal iPSC Reprogramming

Vectors

Safety with confidence

create reproducible ipscs from normal

and diseased cell types with invitrogen

episomal vectors.

CytoTune

_{-iPS Sendai}

Reprogramming Kit

High efficiency for difficult cell

types

reprogram challenging cell types

with the easy-to-use, nonintegrating

cytotune

_{-ips sendai reprogramming}

Which reprogramming technology is right for you?

Well-referenced technology for

iPSC generation from fibroblasts Proven technology for viral-free iPSC generation from normal and diseased cell types

Efficient generation of integration-free iPSCs from multiple somatic cell types

Product name Lentiviral iPSC reprogramming

particles Episomal iPSC Reprogramming Vectors CytoTune

®_{-iPS Sendai}

Reprogramming Kit

cat. no. a1357101, a1357201, a1357301, a1357401, a1357501, a1357601, a1357701, a1357801

a14703 a1378001, a1378002

reprogramming efficiency 0.001–0.01% 0.002–0.08% 0.01–1%

Genomic integration-free no Yes Yes

virus-free reprogramming no Yes no

hands-on time one application, selection and Qc of

putative colonies required one application, minimal Qc of putative colonies required one application, minimal Qc of putative colonies required

blood cell reprogramming no no Yes

convenience hoct4-Gfp and cmv-Gfp reporters available for transduction and expression verification

premixed, 3 plasmid kit; requires neon® _{transfection system or}

similar device

Kit contains all 4 genes required; cannot be purchased separately

price/unit $$ $ $$

available support resources standard protocol for fibroblast

reprogramming protocols for fibroblast reprogramming protocols for fibroblast, cd34+, and pbmc reprogramming; faQs, webinar, publications

$—inexpensive $$—moderate $$$—expensive $$$$—exclusive

Other reprogramming tools

Cells for reprogramming

These cells can be reprogrammed to iPSCs using various

reprogramming methods:

Fibroblasts

• Primary human dermal fibroblasts from either adult or

neonatal samples

Keratinocytes

• Primary human keratinocyte cells from either adult or

neonatal samples

CD34+ cells

• Pooled human hematopoietic progenitor cells derived

from the umbilical cord blood of mixed donors

TaqMan

_{iPSC Sendai Detection Kit}

these taqman

_{probes are used for detecting the presence}

and determining the levels of sendai virus and exogenous

transcription factors (oct3/4, sox2, Klf4, and cmyc) delivered by

the sendai virus from the cytotune

_{-ips sendai reprogramming}

Kit. the primers in the taqman

_{ipsc sendai detection Kit will}

not detect the corresponding endogenous factors.

Cat. No. A13640

Neon

_{Transfection System}

Shockingly simple transfection in stem cells

the neon

_{transfection system, a next-generation}

electroporation technology for highly efficient delivery (~80%)

of nucleic acids such as episomal vectors into somatic cells.

Validation is critical to induced pluripotent stem cell (iPSC)

research. Whether you want to quickly confirm that cells are

pluripotent, or need flexibility in antibody and dye choice,

Molecular Probes

_{labeling products and detection assays}

are an ideal choice. And all can be visualized on the FLoid

Cell Imaging Station. Backed by our extensive internal and

external support resources including the Molecular Probes

Cell Imaging page on Facebook, and the trusted Molecular

Probes

_Handbook.

lifetechnologies.com/detectpsc

FLoid

_{Cell Imaging Station}

Cell imaging right at your benchtop

confidently detect and validate your esc and ipsc

colonies with alkaline phosphatase live stain and

cell surface antibodies on the affordable,

user-friendly floid

_{cell imaging station.}

Cat. No. 4471136

Detection

Alkaline Phosphatase (AP) Live Stain shows robust staining in pluripotent stem cells. H9 human ESC grown on MEF feeders show specific staining of the pluripotent stem cell colonies.

H9 hESCs labeled with pluripotent surface antibodies: SSEA4 Alexa Fluor® _{488 (green), TRA-1-81 Alexa Fluor}® _{594 (red), TRA-1-60 Alexa}

Fluor® _{647 (purple).}

Pluripotent stem cells (PSCs)

Alkaline Phosphatase Live Stain

Quick check for pluripotency

rapidly detect pluripotency in expanding escs and ipscs

without compromising cell integrity with alkaline phosphatase

live stain.

Cat. No. A14353

Antibodies

Flexibility and choice

mix and match your choice of primary antibody and molecular

probes

_{fluorophore for maximum flexibility in pluripotent stem}

cell imaging.

Analysis

Which detection method is right for you?

Allows easy identification of

pluripotency Allows easy identification of pluripotency without compromising cell integrity

Specific and flexible identification of PSCs; antibodies can be combined with different fluorophores for costaining

Product name ELF® _{97 Endogenous}

Phosphatase Detection Kit Alkaline Phosphatase Live Stain Primary antibodies

cat. no. e6601 a14353 lifetechnologies.com/antibodies

specificity medium

(stains stem and progenitor cells) medium (stains stem and progenitor cells) high (stains human escs and ipscs)

terminal assay Yes no no

reaction time stains pscs in 30 min stain pscs typically in 20 min

or less stain pscs in typically 90–120 min color/label Yellow-green fluorescent precipitate

under hoescht/dapi long pass filter Green/fluorescein (fitc) unconjugated primary antibodies, may be detected with secondary conjugated to fluorophore of choice

floid™ _{protocols available no Yes Yes}

unit size 500 µl vial sufficient for staining four 24-well plates, twelve 6-cm dishes, or four 10-cm dishes

50 µl vial sufficient for staining four 24-well plates, twelve 6-cm dishes, or four 10-cm dishes

100 µg vial sufficient for staining four 24-well plates, twelve 6-cm dishes, or four 10-cm dishes

relative price/unit $$ $ $$$

$—inexpensive $$—moderate $$$—expensive $$$$—exclusive

Stem cell research requires cellular and molecular tools to confirm pluripotency of the stem cell population, or the success

of a differentiation experiment. Whether analyzing proliferation, protein levels, gene expression, or epigenetic profiles, Life

Technologies has the right instruments and products for your research. Go to www.lifetechnologies.com/stemcellanalysis

Selected Instruments

Ion Proton

_Sequencer

Genome and exome sequencing on your benchtop

lifetechnologies.com/proton

QuantStudio

_{12K Flex Real-Time PCR System}

The all-in-one qPCR instrument

Delivering our highest throughput, flexibility, and scalability,

the QuantStudio

_{12K Flex Real-Time PCR System features}

5 interchangeable blocks to support 96-well, Fast 96-well,

384-well, TaqMan

_{Array Card, and OpenArray}

_formats.

lifetechnologies.com/quantstudio

Assays

TaqMan

_{Protein Assays}

TaqMan

_{Protein Expression Assays are designed for accurate}

quantitation of protein expression using gold-standard

TaqMan

_{5’ nuclease chemistry with a workflow and}

sample quantity similar to our TaqMan

_{Gene Expression}

and MicroRNA (miRNA) Assays. As the protein expression

results are obtained on the same analytical platform, these

revolutionary assays enable direct correlation of mRNA and/or

miRNA expression to protein expression.

Mesenchymal stem cells (MSCs)

Culture

MesenPRO RS

_Medium

A reduced-serum medium for

MSC culture containing 2% FBS

mesenpro rs

_{medium consistently}

improves expansion of mscs compared

with classical media (dmem + 10% fbs)

and maintains trilineage mesoderm

differentiation potential.

Cat. No. 12746012

Mesenchymal stem cell research

requires different culture conditions

depending on your needs. Life

Technologies provides the broadest

selection of Gibco

_{cGMP compliant}

complete culture systems for

mesenchymal stem cells (MSCs) and

adipose-derived stem cells (ADSCs),

some of which are free of animal-derived

components. These media are designed

to minimize adaptation time, maximize

cell performance, and meet regulatory

requirements. For more information, go

to

lifetechnologies.com/stemcell/msc

Which mesenchymal stem cell culture medium is right for you?

The standard serum-based culture for MSCs and ADSCs Reduced serum medium for lower variability and

cost-savings with serum-culture Fully documented serum-free MSC culture reducing the burden in qualifying reagents, facilitating transition from the bench to the clinic Serum-free, xeno-free MSC culture reducing the risk and variability associated with animal origin components

Product name MSC-Qualified FBS MesenPro RS™ _{Medium StemPro}® _{MSC SFM CTS}™ _StemPro® _{MSC SFM XenoFree}

cat. no. 12662-002, 12662-011, 12662-029 (usda), 12664-025

(australia origin), 12763-025 (us origin) 12746012 e071000 (new Zealand origin) a1033201 a1067501

Recommended applications Multiple species; culture of cells sourced from bone marrow, adipose tissue, cord blood, umbilical cord, placenta

Multiple species; culture of cells sourced from

bone marrow, adipose tissue, umbilical cord Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, hair follicle Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, and cord blood, as well as pericytes and fibroblasts

lot-to-lot variability high medium low low

components/unit 1 x 50 ml, 100 ml, or 500 ml 1 x 500 ml mesenpro rs™ _{basal medium}

1 x 10 ml mesenpro rs™ _{Growth supplement}

1 x 500 ml stempro® _{msc sfm basal medium}

1 x 75 ml stempro® _{sfm supplement}

1 x 500 ml stempro® _{msc sfm basal medium}

1 x 5 ml stempro® _{msc sfm xenofree supplement}

additional components required for complete

medium dmem low glucose Glutamax™_{-i medium}

Glutamax™_{-i medium Glutamax}™_{-i cts}™_{, substrate (cellstart}™ _cts™ _{or attachment factor) Glutamax}™_{-i cts}™_{, substrate (cellstart}™ _cts™ _{or attachment factor)}

supports growth at high cell density; less

medium, surface area, and time no no Yes Yes

relative price/500 ml of complete medium $ $ $$ $$$

stability of complete medium stable at 4°c for 2 weeks stable at 4°c for 15 days stable at 4°c for 4 weeks stable at 4°c for 2 weeks specifically designed for cell therapy

research no no Yes Yes

$—inexpensive $$—moderate $$$—expensive $$$$—exclusive

Human MSCs from a 4-donor, passage 4 pool, expanded for 9 passages in StemPro®

MSC SFM XenoFree on CELLstart™ _CTS™

Substrate–coated flasks.

StemPro

_{MSC Serum Free}

Medium (SFM) CTS

Serum-free medium for growth

and expansion of MSCs

stempro

_{msc sfm cts}

_provides

superior efficiency of human msc

expansion at high cell densities, requiring

less medium, surface area, and time

compared with classical medium (dmem

(low glucose) + 10% fbs) when used

in conjunction with cellstart

_cts

substrate.

Which mesenchymal stem cell culture medium is right for you?

The standard serum-based culture for MSCs and ADSCs Reduced serum medium for lower variability and

cost-savings with serum-culture Fully documented serum-free MSC culture reducing the burden in qualifying reagents, facilitating transition from the bench to the clinic Serum-free, xeno-free MSC culture reducing the risk and variability associated with animal origin components

Product name MSC-Qualified FBS MesenPro RS™ _{Medium StemPro}® _{MSC SFM CTS}™ _StemPro® _{MSC SFM XenoFree}

cat. no. 12662-002, 12662-011, 12662-029 (usda), 12664-025

(australia origin), 12763-025 (us origin) 12746012 e071000 (new Zealand origin) a1033201 a1067501

Recommended applications Multiple species; culture of cells sourced from bone marrow, adipose tissue, cord blood, umbilical cord, placenta

Multiple species; culture of cells sourced from

bone marrow, adipose tissue, umbilical cord Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, hair follicle Human; culture of cells sourced from bone marrow, adipose tissue, umbilical cord, and cord blood, as well as pericytes and fibroblasts

lot-to-lot variability high medium low low

components/unit 1 x 50 ml, 100 ml, or 500 ml 1 x 500 ml mesenpro rs™ _{basal medium}

1 x 10 ml mesenpro rs™ _{Growth supplement}

1 x 500 ml stempro® _{msc sfm basal medium}

1 x 75 ml stempro® _{sfm supplement}

1 x 500 ml stempro® _{msc sfm basal medium}

1 x 5 ml stempro® _{msc sfm xenofree supplement}

additional components required for complete

medium dmem low glucose Glutamax™_{-i medium}

Glutamax™_{-i medium Glutamax}™_{-i cts}™_{, substrate (cellstart}™ _cts™ _{or attachment factor) Glutamax}™_{-i cts}™_{, substrate (cellstart}™ _cts™ _{or attachment factor)}

supports growth at high cell density; less

medium, surface area, and time no no Yes Yes

relative price/500 ml of complete medium $ $ $$ $$$

stability of complete medium stable at 4°c for 2 weeks stable at 4°c for 15 days stable at 4°c for 4 weeks stable at 4°c for 2 weeks specifically designed for cell therapy

research no no Yes Yes

$—inexpensive $$—moderate $$$—expensive $$$$—exclusive

StemPro

_{MSC SFM XenoFree}

Enabling serum-free and

xeno-free expansion of human MSCs

stempro

_{msc sfm xenofree offers}

a completely animal origin–free

system when used in conjunction with

cellstart

_cts

_{substrate; thus,}

mscs and adscs are grown in a more

physiologically similar environment that

allows for more clinically relevant results.

Cat. No. A1067501

StemPro

_LipoMAX

_Supplement

Defined xeno-free lipid

supplement

stempro

_lipomax

_{supplement is a}

human-derived, lipoprotein-based cell

culture supplement for the improved

expansion of mscs and adscs.

Cat. No. A1085001

CELLstart

_CTS

_Substrate

xeno-free, fully defined cell culture

substrate that contains components

only of human origin. cellstart

_cts

substrate enables attachment of human

mscs and adscs.

Mesenchymal stem cells differentiated into fat cells and stained with Oil Red O, a fat-soluble dye.

Fluorescent visualization of StemPro® _{Rat Alk Phos}

Expressing Mesenchymal Stem Cells using ELF® ₉₇

Endogenous Phosphatase Kit.

StemPro

_{Human Adipose-Derived}

Stem Cell (ADSC) Kit

human adscs isolated from human lipoaspirate

tissue and cryopreserved from primary cultures.

Kit contains human adscs and mesenpro rs

medium.

Cat. No. R7788110

Mouse (C57BL⁄6) Mesenchymal Stem Cells

isolated from bone marrow of c57bl⁄6 mouse at

≤8 weeks after gestation.

Cat. No. S1502100

StemPro

_{Rat Alk Phos Expressing}

Mesenchymal Stem Cells

these cells have a unique ability to track cells in

transplantation and differentiation studies.

Cat. No. R7789120

Gibco

_{MSC differentiation kits}

Human MSC differentiation to the adipogenesis,

chondrogenesis, and osteogenesis lineages.

Complement StemPro

_{MSC SFM, MesenPRO}

RS

_{Medium, and MSC-qualified FBS cell}

expansion systems.

StemPro

_{Adipogenesis Differentiation Kit}

complete differentiation into fat cells.

Cat. No. A1007001

StemPro

_{Chondrogenesis}

Differentiation Kit

complete differentiation into cartilage cells.

Cat. No. A1007101

StemPro

_{Osteogenesis Differentiation Kit}

complete differentiation into bone cells.

Cat. No. A1007201

Differentiation

Mesenchymal stem cells (MSCs)

A B C

D E

CD19 Pacific Blue™ _fluorescence

CD14 Pacific Blue™ _{fluorescence HLA DR Pacific Blue}™ _fluorescence

CD45 Pacific Orange™ _{fluorescence FxCycle}™ _{Violet fluorescence}

CD73 Phy coerythrin fluor esc enc e CD73 Phy coerythrin fluor esc enc e Count CD90 FIT C fluor esc enc e CD105 P erCP-Cy ®5.5 fluor esc enc e

Immunophenotype data. Human mesenchymal stem cells collected at passage 7 were identified as: (A) 99.7% CD73+_{/0.29% CD19}–_{; (B) 99.4%}

CD73+_{/0.22% CD45}–_{; (C) 99.7% CD90}+_{/0.09% CD34}–_{; (D) 99.9% CD90}+_{/0.07% CD14}–_{; (E) 99.2% CD105}+_{/0.76% HLA-DR}–_{. Detection was by FACS using}

the Attune® _{Acoustic Focusing Cytometer, and the data demonstrate that cell expression remains as expected after 7 passages.}

Detection

Detection of human mesenchymal stem cells using the Attune

_{Acoustic Focusing Cytometer}

For more information, go to

lifetechnologies.com/attune

The International Society for Cellular Therapy (ISCT) has proposed a set of standards to define hMSCs for laboratory investigations and

preclinical studies, including specific surface antigen expression in which ≥95% of the cells express the antigens recognized by CD105,

CD73, and CD90, with the same cells lacking (≤2% positive) the antigens recognized by CD45, CD34, CD14 or CD11b, CD79a or CD19,

and HLA-DR (Cytotherapy 8:315 (2006)).

Neural stem cells (NSCs)

Culture

Differentiation

B-27

_Supplements

b-27

_{supplements are 50x liquid supplements for growth and}

long-term viability of hippocampal neurons. b-27

_supplement

is the most referenced supplement in neurobiology research,

and is offered in both serum-free and xeno-free versions.

lifetechnologies.com/b27

N-2 Supplement

n-2 supplement is recommended for the growth and

expression of neuroblastomas as well as postmitotic neurons

in primary cultures from both the peripheral nervous system

(pns) and the central nervous system (cns).

Cat. No. 17502048

These supplements are used in combination with a basal

medium such as Neurobasal

_Medium.

StemPro

_{NSC SFM}

serum-free growth and expansion of human neural stem

cells (hnscs), with versatility to support both adherent and

suspension nsc cultures.

Cat. No. A1050901

Neurobasal

_{and Neurobasal}

_{-A media}

serum-free basal media for the long-term viability of

hippocampal and other neurons of the central nervous system.

neurobasal

_{medium is optimized for prenatal and embryonic}

neurons, while neurobasal

_{-a medium is best for growing}

postnatal and adult brain neurons.

lifetechnologies.com/neurobasal

Natural Mouse Laminin

supports the growth of human or rat neural stem cells, as well

as neural primary cells.

Cat. No. 23017015

Human Neural Stem Cells (H9-Derived)

consistent high-purity cells with normal human karyotypes

that can differentiate into neurons, oligodendrocytes, and

astrocytes. consistent proliferation in adherent cell culture

when used with stempro

_{nsc sfm.}

Cat. No. N7800100

Rat Fetal Neural Stem Cells

these cells can be expanded as adherent or neurosphere

suspension cell cultures while more than 75% of nscs retain

undifferentiated phenotype. also supports differentiation into

neurons, oligodendrocytes, and astrocytes.

cat. no. n7744100

Human embryonic stem cells differentiated into neurons.

Gibco

_{media and supplements are the most widely cited reagents for use in neural stem cell culture. Choose between the}

flexibility of our B-27

_{, N-2, StemPro}

_{Neural supplements, and Neurobasal}

_{Medium, or ease of use with the StemPro}

_NSC

SFM kit, specifically formulated for serum-free growth and expansion of human neural stem cells. For more information, go to

Hematopoietic stem cells (HSCs)

Culture

CD34+ cells reprogrammed into induced pluripotent stem cells using the CytoTune®_{-iPS Sendai}

Reprogramming Kit.

Dynal

_{CD34 Progenitor Cell}

Selection System

stem cells can be isolated directly from whole

blood, cord blood, or bone using this positive

magnetic isolation of human cd34+ progenitor

stem cells with bead release.

Cat. No. 11301D

Hematopoietic stem cells expressing CD34 (CD34+ cells) are defined by their ability to replenish all blood cell types and to

self-renew. We offer CD34+ media, cells, and selection tools specifically formulated to support the culture of these stem cells. For

more information, go to

lifetechnologies.com/stemcell/hsc

StemPro

_{-34 SFM}

serum-free medium specifically formulated to

support the development of human hematopoietic

stem cells (cd34+) from sources including bone

marrow, peripheral blood, and neonatal cord

blood. it is manufactured with components of

animal, human, recombinant, or synthetic origin.

Cat. No. 10639011

StemPro

_{CD34+ Cell Kit}

pooled human hematopoietic progenitor cells

(hpcs) derived from the umbilical cord blood

of mixed donors. along with cord blood–derived

cd34+ cells, this kit includes stempro

_{-34 sfm}

basal liquid medium and frozen stempro

_-34

nutrient supplement to facilitate the immediate

culturing of the stempro

_{cd34+ cells.}

Generating unblemished iPSCs

for disease investigation

“If you alter the genetics of the cell type you’re working

with by integrating viruses, you’re going to have an

enormous amount of difficulty teasing out what is the

backbone genetic contribution versus the altered cell

type contribution. By being able to create cells that are

intact and have no modifications to the genome, you can

really feel secure that what you’re looking at now is the

genetic contribution to the disease.

“We were of course thrilled to see that Life Technologies

had made available a new product, the Sendai virus

reprogramming product [CytoTune

_{-iPS Sendai}

Reprogramming Kit], which is as simple as the lentiviral

reprogramming or retroviral reprogramming but has

this major advantage that there are no reprogramming

factors left. It’s fully nonintegrating.

“It makes use of this RNA virus, the Sendai virus,

that’s nonreplicative. You infect the somatic cells just

one time. Usually within 18 to 21 days we see the

appearance of iPS cell colonies. After we’ve expanded

those colonies and actually established a line, by the

time they’ve been passaged between five to eight times

there’s no presence of this RNA virus anymore. So the

lines in essence have an unblemished genome and they

have none of the reprogramming factors left. So you’ve

really got the best possible material for doing the types

of studies we’re interested in.”

Watch the video

at

lifetechnologies.com/ipscvideo

Chad Cowan, PhD,

Assistant Professor

of Stem Cell and

Regenerative Biology at

Harvard University

Introducing LifeLab

™

hands-on

pluripotent stem cell workshops

Life Technologies is proud to announce the opening of LifeLab

_{Stem Cell Research Centers in Carlsbad, CA, and}

Madison, WI. These centers provide customers with hands-on training for culturing and characterizing human

embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), as well as reprogramming techniques for

the creation of iPSCs.

For research use only. Not intended for any animal or human therapeutic or diagnostic use. ©2012 Life Technologies Corporation. All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. Accutase is a registered trademark of Innovative Cell Technologies. CytoTune is a registered trademark of DNAVEC Corporation. TaqMan® _{is a registered trademark of Roche Molecular Systems,}

Inc., used under permission and license. B-27 is a registered trademark of Southern Illinois University. Cy is a registered trademark of GE Healthcare. Facebook is a trademark of Facebook, Inc. CO25406 0512

For more information, go to

lifetechnologies.com/stemcell

publications—protocols—_videos —

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