Diabetes Research and Treatment

Open Access

Review

http://dx.doi.org/10.14437/DRTOA-1-102

Received Date: July 10, 2014

Accepted date: August 23, 2014
Published date: August 26, 2014

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Encapsulation of Insulin Producing Cells for Diabetes Treatment Using

Alginate and Cellulose Sulphate as Bioencapsulation Polymers

Brian Salmons1, Eva Maria Brandtner1,2, John A Dangerfield1 and Walter H Gunzburg1,3
1

Austrianova Singapore, 20 Biopolis Way, #05-518 Centros, Singapore

Vorarlberg Institute for Vascular Investigation and Treatment (VIVIT), LKH Feldkirch, Carinagasse 47, A-6807 Feldkirch, Austria

Department for Pathobiology, Institute of Virology, University of Veterinary Medicine, Veterinaerplatz 1, A-1210 Vienna, Austria

!"#$%&'()* Diabetes; Diabetes type 1; Diabetes type 2;

!"#$%&'$(

Hypoglycaemia; Normoglycaemia;

Cell encapsulation;

It has long been the holy grail of cell encapsulation to treat

alginate; Cellulose sulphate; Clinical trials*
diabetes with insulin producing cells that are shielded from

+,,&"-./0.%1()* pDADMAC

Polydiallyldimethyl

rejection or clearance by the host immune system. Indeed,

ammonium chloride, Poly-L-Lysine (PLL)*
the idea has been around for more than 50 years with the
first demonstration in an animal model being published in
1980 and the first patient treated being reported in 1994. In

Introduction
the meantime, more clinical experience has been accrued
Worldwide, in 2013, an estimated 382 million people
with the use of encapsulated cells for the treatment of
had diabetes and more than 30 million of these people suffer
human diabetes. Most of these studies have focused on the
from type 1 diabetes (also known as insulin dependent
use of alginate as the encapsulation material and although
diabetes or juvenile diabetes) which results from the
some promising data has been obtained and is reviewed in
autoimmune mediated death or destruction of insulinthis article, it is still far from being a product that can be
producing beta cells that are located in the islets of
routinely

applied.

Cellulose

sulphate

has

excellent
Langerhans within the pancreas [1,2]. Typically, type 1

properties making it ideal for cell encapsulation and has

diabetes is treated with insulin replacement therapy delivered
been shown to be safe in 27 patients. Moreover, data has
either via subcutaneous injection or by an insulin pump and it
been obtained showing safety and efficacy in a rat model of
usually requires at least three or more daily insulin injections,
diabetes supporting that cellulose sulphate might be the
which is cumbersome. Insulin administration is accompanied
material of choice for this application.
by the need for dietary management and this typically includes
carbohydrate tracking as well as careful monitoring of blood

23%&&"(4%1'.15* +607%&)

Walter

Department

Institute

Gunzburg,
glucose levels using glucose meters.

for

Pathobiology,

of

Virology,

University of Veterinary Medicine, Veterinaerplatz 1, A1210 Vienna, Austria

Copyright: 2014 DRTOA. This is an open-access article distributed under the terms of the Creative Commons Attribution License, Version 3.0, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Volume 1 Issue 1 102

www.aperito.org

Citation: #SJBO
4BMNPOT
(2014),

Encapsulation of Insulin Producing Cells for Diabetes Treatment Using Alginate and Cellulose Sulphate as

Bioencapsulation Polymers. DRTOA 1: 102.

Page 2 of 7

http://dx.doi.org/10.14437/DRTOA-1-102

The most common insulins that are currently used for

levels fall then insulin production is down regulated i.e.

treatment are biosynthetic products produced using genetic

giving physiological control of insulin production (Figure 1).

recombination techniques [2, 3]. Nevertheless, hypoglycaemic

events can occur and these episodes can lead to life
threatening incidents as well as long term and ongoing organ
damage. The transplantation of beta cells or islets is an
alternative treatment that allows insulin to be produced from
replacement cells, mimicking the normal physiology much
more closely than exogenous insulin injections. This kind of
approach leads to a reduction in the incidence of
hypoglycemia, improved glycemic control and overall
improvement in quality of life [2, 3]. Moreover, beta-cell
replacement therapy should also provide great benefit for
some people with type 2 diabetes mellitus, where beta-cell
insufficiency plays a central role in its pathogenesis [3].

Although initial attempts at islet transplantation showed the

potential successes of such a strategy for the treatment of

Figure 1: (A) Schematic representation of encapsulated cells

diabetes, there are a number of problems that limit the use of

and the principle behind the immunoprotection of such cells.

this treatment to relatively few patients due to (i) the need for

Small molecules such as nutrients, blood sugar etc can freely

at least two donor pancreases to treat each time due to poor

enter the capsule and reach the cells inside so that cells can

vascularization and thus hypoxia, patient (ii) graft failure,

survive and even divide. Likewise, products produced from

which occurs within a relatively short period of destruction by

the cells such as insulin can be released from the encapsulated

autoimmunity and allorejection, as well as the toxic effects of

cells. Larger molecules and cells of the immune system cannot

immunosuppressive

for

enter the capsule thus foreign cells cannot be rejected or

immunosuppression results in vulnerabiility to infections as

destroyed (B) Electron microscopic image of an intact

well as to various forms of cancer [2, 3].

cellulose sulphate capsule. (C) Electron microscopic image of

drugs.

Moreover,

the

need

a freeze fractured cellulose sulphate capsule. Visible are single

Cell Encapsulation for the Treatment of Diabetes

cells within the capsule (red arrow). Reference bar in panel B
Many researchers have turned to immunosuppressive devices
and C represents 100 m.
as a means to protect transplanted beta cells so that they
survive longer in the patient resulting in fewer cells being

Source of cells

required. However, it should be noted that, at present, the

In theory a number of different cell sources could be used for

minimal number of cells required for therapeutic benefit is still

encapsulation for the production of insulin including human

controversial. Nevertheless, since these immunoprotective

islets or beta cells as well as those from other species. Most

devices are porous, the cells can respond physiologically to

often, porcine islets have been the species of choice even

high blood sugar by producing insulin but when blood sugar

though there are concerns about adventitious agents [4].

Nevertheless, at least most known adventitious agents can be

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Citation: Walter H Guzburg (2014),

Encapsulation of Insulin Producing Cells for Diabetes Treatment Using Alginate and Cellulose Sulphate as

Bioencapsulation Polymers. DRTOA 1: 102.

Page 3 of 7

http://dx.doi.org/10.14437/DRTOA-1-102

avoided and cells prescreened [5]. Also as long as the cells

when it is not properly bound to alginate and it may decrease

remain in the capsule, agents such as viruses might not be

encapsulated cell viability if used in too high concentrations

released (although there are reports that at least retroviruses

[12, 13]. Recently improvements have been reported that

can be released from encapsulated cells [6]. Other sources of

appear to have addressed some of the issues with the use of

cells include human stem cells that can be differentiated into

alginate [14].

insulin producing cells such as mesenchymal stem cells [7] as

well as cells that are genetically modified to produce insulin

Cells encapsulated in alginate have been used in clinical trials

[8].There has been some concern that certain earlier clinical

for diabetes. The results from the first type 1 diabetic patient

studies may have been undertaken too soon and the

to be treated with alginate encapsulated human islets was

International Xenotransplantation Association has published a

reported 20 years ago and showed insulin independence and

consensus statement on conditions for undertaking clinical

tight glycaemic control for 9 months after the intraperitoneal

trials of porcine islet products in type 1 diabetes that covers

injection of the encapsulated capsules islets [15]. However, it

preclinical efficacy and complication data required to justify a

should be noted that this patient had previously received a

clinical trial [9].

kidney graft and was thus still under immunosuppression so

the immunoprotective properties of the encapsulation device

Encapsulation materials
was not rigorously tested in this study.
There are a number of different devices that are being
developed to house beta cells or islets for the treatment of

In the last 10 years later, there have been a number of clinical

diabetes. Lim and Sun first reported in 1980 successful

studies with similar outcomes and that demonstrate some

reversal of diabetes in rat recipients of immunoisolated islets

degree of safety and bio-invisibility (i.e. absence of a wide

for up to 15 weeks [10]. They encapsulated islets in the algae

array of islet cell-directed as well as anti- MHC class III

derived polymer, alginate, with Poly-L-Lysine (PLL) and

antibodies) but only partial and transient metabolic benefits

polyethylenimine coatings. Although alginate continues to be

have been observed in these studies [16-20]. C-peptide could

a popular choice for cell encapsulation, other materials such as

be detected in the first few days after transplantation of the

Cellulose Sulphate (CS)(see below), agarose, chitosan,

alginate encapsulated cells but standard insulin therapy

polyhydroxyethylmetacrylate-methyl-methacrylate,

eventually had to be resumed. A recent clinical study with

copolymers of acrylonitrile and polyethylene glycol are

alginate encapsulated porcine islets cells have shown that

available as well [11]. It is also common to coat alginate

although insulin is still required, both the insulin dose and the

with a polycation, which binds to negatively charged alginate

episodes of unaware hypoglycaemia can be reduced [21].

molecules.
The most commonly used polycations are PLL as well as poly-

In some of the trials, there was evidence that the alginate

L-ornithine, poly-D-lysine and polymethylene-co-guanidine.

capsules induced a strong foreign body reaction and the

This procedure has several advantages, such as increasing the

capsules were overgrown by fibrous connective tissue

mechanical stability of the microcapsule, allowing the control

(pericapsular fibrotic overgrowth) that may have affected the

of the membrane porosity and preventing the beads from being

ability to produce insulin or even the survival of the cells once

dissolved by physiological chelating agents. However, soluble

transplanted [12]. Various factors have been identified as

and non-complexed PLL has been shown to be an

playing a role in the suboptimal biocompatibility of alginate

inflammatory molecule, which leads to fibrotic overgrowth

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Citation: Walter H Guzburg (2014),

Encapsulation of Insulin Producing Cells for Diabetes Treatment Using Alginate and Cellulose Sulphate as

Bioencapsulation Polymers. DRTOA 1: 102.

Page 4 of 7

http://dx.doi.org/10.14437/DRTOA-1-102

capsules [22-27] including induction of cytokines that

Good biocompatibility for the cells in the microcapsule.

stimulate human monocytes [23] and also the presence of

Many different cell types and cell lines have been

protein contaminants [24].

successfully encapsulated e.g. HEK293, CHO cells, betacells and pancreatic islets, fibroblasts, epithelial cells,

Although there have been many attempts to improve the

hybridomas and a variety of different stem cells from

biocompatibity of alginate capsules, for example by improving

various sources [38]

the purity of the starting materials [28] and improving the

Encapsulated cells are able to grow and survive for

mechanical strength of alginate capsules, regardless of the

extended periods in the capsule but three-dimensional

type of cell that is encapsulated [14], clearly there are still a

contact inhibition prevents the release of cells from the

number of problems associated with the use of alginate for cell

capsules.

encapsulation [12].

Good biocompatibility and inertness of the capsules when

implanted at various sites in the body.

Sodium cellulose sulphate is ideal as a cell encapsulation

Lack of an immune or inflammatory response either to the

material [29]. The cellulose sulphate encapsulation technology

cellulose sulphate starting solution, the capsule material or

was originally developed for the treatment of solid tumours

to the cells that are protected by it.

such as pancreatic and breast cancer [30-32]. Encapsulated

cells were tested in a variety of preclinical animal models as

Demonstrated to be safe and well tolerated in three clinical

trials and numerous animal studies [30-32, 38].

well as in clinical trials and shown to be efficacious.

Lack of fibrous overgrowth of the implanted capsules.
Moreover, the implanted encapsulated cells have a well
Large scale GMP manufacturing of an encapsulated cell
documented, long term, safety profile in a total of 27 patients medicinal product has been achieved [38].
even when they remained in the body for up to two years [30 Ability to freeze the encapsulated cells for long term
33].
storage as well as facilitating shipment around the
As we have previously noted [29], capsules consisting of
world.
polymers of cellulose sulphate and polydiallyldimethyl
ammonium chloride (PDADMAC) (Figure 1) offer a number

Both insulin producing cell types showed good viability upon

of advantages as well as unique properties (Table 1). The

encapsulation and respond to increasing concentrations of

cellulose sulphate cell encapsulation technology has been used

glucose by producing insulin in a dose response fashion.

to encapsulate many different cells types, including insulin

Indeed, in the later study, statistical analysis revealed no

producing cells such as primary porcine islet cells [34] and

differences in glucose-dependent cell proliferation, insulin

hamster beta (HIT-15) cells [35].

secretion and glucose uptake between non encapsulated and

encapsulated HIT-T15 cells and stimulation of HIT-T15 cells

Table 1 some advantages of cellulose sulphate

with 100 mg/ml glucose resulted in an insulin secretion

for living cell encapsulation

response that was biphasic [35].

Ability to reproducibly source, produce and characterize the

More recently, cellulose sulphate encapsulated porcine islets
starting material.
were tested in immune competent Sprague Dawley rats made
Robustness and flexibility of the capsules (permitting
diabetic by intraperitoneal injection of streptotocin. Blood
delivery through needles or catheters without bursting).
sugar levels were monitored on a weekly basis. Porcine

Volume 1 Issue 1 102

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Citation: Walter H Guzburg (2014),

Encapsulation of Insulin Producing Cells for Diabetes Treatment Using Alginate and Cellulose Sulphate as

Bioencapsulation Polymers. DRTOA 1: 102.

Page 5 of 7

http://dx.doi.org/10.14437/DRTOA-1-102

C-peptide levels and insulin levels were measured using

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Encapsulation of Insulin Producing Cells for Diabetes Treatment Using Alginate and Cellulose Sulphate as

Bioencapsulation Polymers. DRTOA 1: 102.

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Bioencapsulation Polymers. DRTOA 1: 102.

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