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AAAS/Science Business Office Feature

Cell therapy has technically been around for de-

cades—think bone marrow transplants—but now

researchers are fi nding evermore sophisticated

ways to manipulate and use cells for medicine.

Physicians and scientists are harvesting dendritic

cells, T cells, stem cells, and more from patients’ bodies and

identifying and separating those with the strongest potential

for helping patients. Once isolated, these cells can be grown

in large quantity and returned to a patient’s body to treat vari-

ous diseases.

Though cell therapy remains a lively area of research,

commercially available treatments have been slow to enter

the clinic. The U.S. Food and Drug Administration (FDA)

approved the fi rst-ever autologous cell therapy (which ma-

nipulates the patient’s own cells and returns them to his or

her body), Genzyme’s Carticel, for treating damaged knee

cartilage in 2007. The second, Provenge, for treating meta-

static prostate cancer, followed in 2010. Last March, the

Centers for Medicare and Medicaid Services announced

that it plans to cover this $93,000 prostate treatment, a ma-

jor boost for Dendreon, the Seattle-based company that

makes the drug.

Allogeneic cell therapies, made from cells harvested from

donors rather than the patient, offer the possibility of “off the

shelf” use. Dozens of candidates are currently in clinical tri-

als for applications ranging from boosting wound healing to

treating graft-versus-host disease to supplementing blood

cancer chemotherapy treatments.

Because these cells are being used for medicinal purposes,

the FDA regulates the entire process under the same rules

that govern the production of pharmaceuticals, known as

good manufacturing practice (GMP). Given the vast potential

for cell therapy, many companies making instruments, re-

agents, and other tools for cell separation have shifted their

SORTING CELLS FOR MEDICINEThe basic technology for cell separation hasn’t changed

much over the past several years. What has changed

dramatically is what those cells are being used for:

treating patients. As the promise of cell therapy is

gradually realized, researchers are searching for cell

separation technologies that can solve some basic

problems, such as improving cell processing speed

and making processing as sterile as possible. Research-

ers already use a mix of density-gradient separation,

magnetic separation, and fl ow cytometry to prep cells for

clinical use but hope for newer chip-based technologies to

help advance the fi eld. By Anne Harding

“Given the rigid

regulatory

environment, the

manufacturers in

this clinical space

need to have a

long-term outlook

and a commitment

to cellular therapy and

regenerative medicine.”

focus away from the research lab and into the translational

and clinical research space.

But researchers in the fi eld say many gaps remain in what’s

available to develop clinically viable cell therapies, from in-

struments to antibodies. And because the payoffs so far

have been relatively few and far between, some companies

making these instruments or reagents have had to pull their

products off the market, such as with Baxter’s Isolex 300

Magnetic Cell Selection System—leaving researchers who

use their products high and dry, especially those with open

clinical trials under way.

“It’s really hard to depend on a single manufacturer,” says

Lynn O’Donnell, who directs the Cell Therapy Laboratory at

the Ohio State University’s James Cancer Hospital in Co-

lumbus. “Given the rigid regulatory environment, the manu-

facturers in this clinical space need to have a long-term out-

look and a commitment to cellular therapy and regenerative

medicine.”

BACK TO BASICS: CELLS OUT OF TISSUESeparating cells from their tissue of origin is one of the real

bottlenecks for developing cell therapies for clinical use,

says Firman Ghouze, director for cell therapy within GE

Healthcare Life Sciences. While sorting cells out of blood

is fairly easy, isolating them from fat—which has turned out

to be a very rich source of stem cells—is another matter.

LIFE SCIENCE TECHNOLOGIES

CELL SEPARATION TECHNOLOGIES

AAAS/Science Business Office Feature

www.sciencemag.org/products 737

Companies are putting a lot

of effort towards develop-

ing simpler high volume,

high throughput processes

for separating cells from

tissue that can be handled

by technicians and not

only Ph.D.s, according to

Ghouze.

To this end, GE offers the

StemSource 900/MB Tissue

Processing System, which

was developed by Cytori

Therapeutics. This sterile,

closed centrifugation sys-

tem can be used for extract-

ing several different regenerative cell types from connective

tissue, yielding an average of 4.6 x 107 nucleated cells per 100

grams of tissue. GE’s Res-Q 60 Bone Marrow Concentration

System, also centrifuge-based, can be used at the patient’s

bedside to isolate mononuclear cells from bone marrow in

less than 20 minutes.

O’Donnell and her colleagues separate islet cells out of

pancreatic tissue using another popular centrifuge-based

instrument, CaridianBCT’s COBE 2991 Cell Processor. The

instrument uses a closed fl uid path and single-use dispos-

able set for cell centrifugation—a vast improvement over

traditional centrifuges and conical tubes because it allows

various fractions to be collected within a closed system,

O’Donnell explains.

But the density gradient system has limitations when it

comes to coping with pancreatic islets, which “are kind

of like mini-organs because they are clusters of cells,” she

adds. The instrument is nonspecifi c and variable between

islet isolations.

“It’s challenging to come up with a purifi cation technique

that works for large clusters of cells and variably sized cells,”

O’Donnell says. And given the nature of islets, “you end up

sacrifi cing yield in order to gain purity because the separation

isn’t great.”

CLOSED SYSTEMS ARE KEY FOR CLINICAL PRODUCTSMost researchers in the fi eld, including O’Donnell, use a

combination of centrifugation, magnetic bead separation,

and fl ow cytometry to develop their cell therapies. Magnetic

separation has the advantage of speed, while cell sorting al-

lows for the use of multiple markers and results in a purer

end product.

“In the case of magnetic bead systems, a whole new se-

ries of reagents and equipment have been developed that

allow for more automated processing and more closed oper-

ation, so you can get a sterile cell product,” says Shelly Heim-

feld, who directs the Cellular Therapy Laboratory and cGMP

Cell Processing Facility at the Fred Hutchinson Cancer

Research Center in Seattle. Heimfeld is also past president

of the International Soci-

ety for Cellular Therapy, an

organization that supports

the translation of cell thera-

py research into clinical use.

Such instruments include

Miltenyi Biotec’s Clini-

MACS platform, which uses

magnetic-based technology

for enriching target cells or

depleting unwanted cells.

Miltenyi is about to introduce

a second-generation version

of the instrument, the Clini-

MACS Prodigy. “The idea

is to have the entire GMP-

conformed process for cell manufacturing integrated into one

device,” explains Jurgen Schmitz, head of research and devel-

opment at the Bergisch-Gladbach, Germany-based company.

This closed system can wash, separate (with density gradi-

ent cell separation or magnetic bead cell separation), and for-

mulate the cells of interest, Schmitz explains.

“That is something quite complex in terms of logistics,” he

adds. “Such an instrument can really help make things eas-

ier and less expensive, for example by reducing clean-room

requirements.”

Because companies are focused on making cell-sorting in-

strumentation and reagents that are suitable for use in clinical

products, they are now “thinking about how to comply with

GMP-manufacturing guidelines,” says Tim Fong, technical di-

rector of cell therapy at BD Biosciences.

Maintaining the sterility of cell products is key. BD Biosci-

ences, for example, offers a replaceable gamma-irradiated

fl uidics kit for the Infl ux cell-sorting platform. “All the tubing

that the cell sample comes into contact with on the instru-

ment is single use and gamma irradiated, which mitigates the

prospect of serial contamination,” explains Jack Dunne, who

leads the company’s cell-therapy research team.

ACCELERATING CELL SORTING WITH MULTI-MICROFLUIDICSThe ideal technology for producing cell therapeutics, says

O’Donnell, would be a speedier version of fl ow cytometry

since “you can look at multiple markers and purify very spe-

cifi c kinds of cells.” But, she points out, there’s the challenge

of engineering a single-use fl uid path that allows cell sorting

under sterile conditions. One solution, she says, is to put a

box around a standard fl ow sorter that would provide clean,

HEPA-fi ltered air, “but you still have cross-contamination

questions, like what do you do between processing for pa-

tient A and patient B.”

Another possibility researchers foresee is the use of mi-

crofl uidics chips that sort cells via microscopic channels

and gates that open and close to determine the cells’ fl ow

path. With several fl ow paths running in parallel on a single

chip, the sorting process could be sped up

“In the case of magnetic bead systems, a whole

new series of reagents and equipment have

been developed that allow for more automated

processing and more closed operation, so you

can get a sterile cell product.”

CR

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continued »

LIFE SCIENCE TECHNOLOGIES AAAS/Science Business Office Feature

CELL SEPARATION TECHNOLOGIES

www.sciencemag.org/products738

donor and recipient to pre-

vent rejection. “We used to

think that these [T cells] could

only be used in essentially

closely matched tissue recipi-

ents, but it appears they can

be used across some tissue

types,” he explained.

LOTS OF PROMISE, BUT SLOW PROGRESS TO THE CLINIC“There’s a lot of things going

on” in the development of cellu-

lar therapies, but progress is

“slow and painful,” says Rob-

ert S. Negrin, a professor of

medicine at Stanford Uni-

versity, director of the uni-

versity’s Bone Marrow Trans-

plant Program, and medical

director for the Stanford Cell

Therapeutics Laboratory.

One factor delaying things,

explains Negrin, is the scar-

city of antibodies that are

made to GMP standards for

magnetic cell separation. He

estimates that there are cur-

rently a dozen or so antibod-

ies, made by BD, Miltenyi,

and other companies, that are suitable for preparing cells for

clinical use. “That’s where we often get stuck, is having the

proper reagents,” Negrin says. Just having more antibodies

available for clinical use could give the fi eld a major push for-

ward, he adds. Yet, as with many other technologies related

to the fi eld, investors and companies are shying away from

such investments until they see some real success.

However, investments in products that support cell therapy

research is just as essential as the research itself, says DiGi-

usto, since “one without the other does no good.”

Despite the challenges of developing commercially avail-

able cell therapies, more than a dozen companies have al-

logeneic or autologous products in various stages of clinical

trials, including Geron, Osiris Therapeutics, Cytori Therapeu-

tics, and Aastrom Biosciences.

And with each success story, such as Dendreon’s prostate

cancer drug therapy, Provenge, the fi eld as a whole benefi ts,

explains DiGiusto. “It’s at least a sign to the investment com-

munity that [success] can happen,” he adds. “And it’s really

a critical thing that the investment community continues to

have confi dence in the fi eld.”

DOI: 10.1126/science.opms.p1100055

Anne Harding is a freelance science writer based near New York City.

considerably. What’s more,

the system could be entirely

closed and aseptic. Feasibil-

ity studies by developers,

including Owl Biomedical,

have been promising, ac-

cording to David DiGiusto,

who directs the Labora-

tory for Cellular Medicine

at the Beckman Research

Institute of the City of Hope

in Duarte, California.

Yet another advantage of

electrostatic cell sorting on

chips is that it would be “la-

bel-free.” While centrifuga-

tion techniques are label-free

also, both magnetic cell sepa-

ration and fl ow cytometry

require tagging cells of inter-

est with antibodies or dyes,

creating another challenge

for researchers: fi nding ways

to remove the labels such as

using enzymes that can “clip

off” antibodies after sorting

or designing biodegradable

labels that simply fall apart.

“We’re still in an era where

we’re using the [same] meth-

ods that identify the cells

to mediate their separation,” says Adrian Gee, a professor

in the Department of Cell and Gene Therapy at the Texas

Children’s Hospital.

EVADING REJECTION TO MAKE ‘OFF-THE-SHELF’ PRODUCTSSo far, large pharmaceutical companies seem to have held

back from getting involved in cell therapeutics, which remains

pretty much the province of startup biotechs and translational

research institutions.

“It’s much more attractive for companies to have a gen-

eralized cell product that could be used to treat a wide vari-

ety of patients,” says Gee. But developing such generalized

products requires fi guring out a way to ensure that the recipi-

ent’s immune system accepts the donated cells. Just as with

blood transfusions and organ donations, cell therapies can

be rejected by a patient’s body. Solving this problem may, in

some cases, require using tissue matching techniques like

those used for blood donation or organ transplantation.

Investigators have also learned that some cells, such as

mesenchymal stromal cells and nerve cells, do not provoke

the rejection responses seen with other cell types. Gee and

his colleagues found that T cells primed to attack viruses

can circumvent human leukocyte antigen (HLA) barriers, the

immune-system “tissue types” that must match between

v

Aastrom Bioscienceswww.aastrom.com

Baxterwww.baxter.com

BD Bioscienceswww.bdbiosciences.com

CaridianBCTwww.caridianbct.com

Centers for Medicare and Medicaid Serviceswww.cms.gov

City of Hope’s Beckman Research Institutewww.cityofhope.org/research/beckman-research-institute

Cytori Therapeuticswww.cytori.com

Dendreonwww.dendreon.com

Fred Hutchinson Cancer Research Centerwww.fhcrc.org

GE Healthcare Life Scienceswww.gelifesciences.com

Genzymewww.genzyme.com

Geronwww.geron.com

International Society for Cellular Therapywww.celltherapysociety.org

Miltenyi Biotecwww.miltenyibiotec.com

Ohio State University’s James Cancer Hospitalcancer.osu.edu

Osiris Therapeuticswww.osiristx.com

Owl Biomedicalwww.owlbiomedical.com

Stanford Universitywww.stanford.edu

Texas Children’s Hospitalwww.texaschildrens.org

U.S. Food and Drug Administrationwww.fda.gov

FEATURED PARTICIPANTS

AAAS/Science Business Office FeatureLIFE SCIENCE TECHNOLOGIES

CELL SEPARATION TECHNOLOGIES

AAAS/Science Business Office Feature

www.sciencemag.org/products 739

Newly offered instrumentation, apparatus, and laboratory materials of interest to researchers in all disciplines in academic, industrial, and governmental organizations are

featured in this space. Emphasis is given to purpose, chief characteristics, and availability of products and materials. Endorsement by Science or AAAS of any products or

materials mentioned is not implied. Additional information may be obtained from the manufacturer or supplier.

Electronically submit your new product description or product literature information! Go to www.sciencemag.org/products/newproducts.dtl for more information.

PLATE WASHER/COATER

SQUIRT can wash any 96-, 384-, and 1536-well SBS formatted

plate without manifold exchange or adjustment. SQUIRT’s unique

design sweeps a blade of liquid across the whole surface of the

plate. This is followed by an air blade to dry the plate. This unique

blade design eliminates clogging and the ‘corona effect’ (where

cells are selectively removed from the center of the wells with tra-

ditional probe based instruments). SQUIRT microplate washers are

perfect for any lab performing cell-based assays, enzyme-linked

immunosorbent assays (ELISA), coating microplates, high content

screening, and proteomics and genomics research. The SQUIRT

can be fi tted with a magnetic plate insert for applications using

magnetic beads.

Matrical Bioscience

For info: 509-343-6225 www.matrical.com

EX-VIVO CELL-THERAPY EXPANSION

The Cellbase CT system, which was developed in collaboration

with cell-therapy experts, can be used in a good manufactur-

ing practice (GMP) facility for rapid scale-up of existing manual

cell-culture processes in T fl asks. Cellbase CT provides research-

ers with a fully contained, aseptic cell-expansion area where the

risk of external contamination or internal cross-contamination has

been eliminated. This enables scientists to manufacture multiple

autologous cell therapies from many individual patients in paral-

lel, making the Cellbase CT a cost-effective alternative to labor-

intensive manual cell culture. Cellbase CT comes with a range of

tools to support validation in a GMP-regulated environment and

has been developed and extensively tested to enable clean, re-

producible expansion of clinically applicable stem cells and other

cell types.

TAP Biosystems

For info: 302-478-9060 www.tapbiosystems.com

MAGNETIC BEADS

Streptavidin Mag Sepharose magnetic beads have a high binding capacity for effi cient

purifi cation of biotinylated biomolecules and enrichment of target proteins. The mag-

netic beads provide simplifi ed handling, as rapid capture is facilitated by magnetic de-

vices while delivering scalability and high yield, from low microliter- to high milliliter-scale

sample volumes. Streptavidin Mag Sepharose beads support reliable sample preparation

for downstream analysis and have more than a 400-fold enrichment factor to enable an

increased identifi cation rate of target proteins. The magnetic beads are hydrophilic with a

high density to ensure the beads do not aggregate, avoiding sample loss and increasing

yield. Additionally, the MagRack Maxi magnetic rack provides easy handling and sup-

ports sample capacities of up to 50 ml, to provide capture of low-expressed target pro-

teins from larger sample volumes. The MagRack Maxi consists of an anodized aluminium

housing with a detachable plastic bar containing a neodymium magnet.

GE Healthcare

For info: 800-526-3593 www.gelifesciences.com

MONOCLONAL ANTIBODIES

Monoclonal antibodies have become invaluable tools for research-

ers developing diagnostic tests and novel therapeutics. The 1,500

new mouse monoclonal TrueMAB antibodies have been specifi cal-

ly developed with full-length human proteins produced in HEK293T

cells and validated for critical research and diagnostic needs that

require improved antibody sensitivity and specifi city. TrueMAB an-

tibodies and hybridoma cell lines are generated using OriGene’s

large collection of full-length human proteins as primary antigens

that have been affi nity purifi ed under native conditions to preserve

natural protein conformations. As a result, TrueMAB antibodies pro-

vide high sensitivity and specifi city for the recognition of native epit-

opes on the protein’s natural conformational structure. All TrueMAB

antibodies are routinely validated for critical applications such as im-

munohistochemistry, immunofl uorescent staining, fl ow cytometry,

immunoprecipitation, and Western blot analysis.

OriGene Technologies

For info: 888-267-4436 www.origene.com

AUTOMATED CELL EXPANSION

The Quantum Cell Expansion System is designed to automate cell

culture in a closed system. The cutting-edge technology is intended

to support advancements in the fi eld of cell therapy by streamlin-

ing cell culture processes while reducing the risk of contamination

for large-scale cell manufacturing. Current methods used to culture

cells are manual and complex, which can limit the ability to pro-

vide cellular therapy to a large number of patients. As an integrat-

ed, closed system, the Quantum Cell Expansion System improves

the effi ciency of the cell culture process—allowing for larger scale

manufacturing of cells under good manufacturing practice (GMP)

process control and with less risk of contamination.

CaridianBCT

For info: 877-339-4228 www.caridianbct.com

LIFE SCIENCE TECHNOLOGIES AAAS/Science Business Office Feature

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