Cells can secrete biotherapeutic molecules that can replace or restore host function. The transplantation of such cells is a promising therapeutic modality for the treatment of several diseases including type 1 diabetes mellitus. These cellular grafts are encapsulated in semipermeable and immunoisolative membranes to protect them from the host immune system, while allowing the transport of nutrients and small molecules that are required for cell survival and function. The authors report on SU-8-based biocompatible immunoisolative cuboid microcontainers for cell transplantation. Each microcontainer comprises a 300×300×250 or a 1100×1100×250 μm 3 SU-8 hollowed cuboid base that houses the cells and an optically transparent SU-8-based nanoporous lid that closes the device. The hollowed cuboid base was formed by conventional optical lithography to have 8 nl (200×200×200 μm 3 ) encapsulation volume for cellular payload. The lid comprises a thick SU-8 slab with an array of cylindrical wells, whose bottom surface is sealed with a thin nanoporous SU-8 membrane. The nanoporous membrane was created from a 100 nm grating (width and spacing) initial silicon mold subjected to a repeated cycle of oxidation and wet etching to achieve a 20 nm wide and 200 nm pitch nano silicon grating. Nanoimprinting and oblique-angle metal deposition, followed by inductively coupled plasma etching were utilized to create 15 nm wide and 350-450 nm deep nanoslots in the thin SU-8 membrane. Isolated mouse islets were encapsulated in the hollowed cuboid base and the nanoporous lid was assembled on top. The penetration of large and small molecules into the microcontainer was observed with fluorescence. a)barjorg@yahoo.com.

With strides in stem cell biology, cell engineering and molecular therapy, the transplantation of cells to produce therapeutic molecules endogenously is an attractive and achievable alternative to the use of exogenous drugs. The encapsulation of such cell transplants in semi-permeable, nanoporous constructs is often required to protect them from immune attack and to prevent their proliferation in the host. However, effective graft immunoisolation has been mostly elusive owing to the absence of a high-throughput method to create precisely controlled, high-aspect-ratio nanopores. To address the clinical need for effective cell encapsulation and immunoisolation, we devised a biocompatible cellencapsulating microcontainer and a method to create highly anisotropic nanopores in the microcontainer's surface. To evaluate the efficacy of these nanopores in oxygenating the encapsulated cells, we engineered 9L rat glioma cells to bioluminesce under hypoxic conditions. The methods described above should aid in evaluating the long term survival and efficacy of cellular grafts.

Abbreviations: (Cu) copper, (E) electron, (FIB) focused ion beam, (KRH) Krebs-Ringer solution, (MEMS) microelectromechanical system, (MRI) magnetic resonance imaging, (MRM) magnetic resonance microscopy, (RF) radio frequency, (TEA) tetraethylammonium Keywords: cell encapsulation, cell encapsulation therapy, cell therapy, immunoisolation, immunoprotection, microcontainers AbstractPresent-day islet encapsulation techniques such as polymer microcapsules and microelectromechanical system (MEMS)-based biocapsules have shown promise in insulin replacement therapy, but they each have limitationsthe permeability characteristics of existing polymeric capsules cannot be strictly controlled because of tortuosity and the large size of present-day MEMS biocapsules leads to necrotic regions within the encapsulation volume. We report on a new microcontainer to encapsulate and immunoprotect islets/β cells that may be used for allo-or xenotransplantation in cell-based therapy. The microcontainers have membranes containing nanoslots to permit the bidirectional transport of nutrients, secretagogues, and cellular products while immunoprotecting the encapsulated cells. The 300-µm microcontainers were fabricated from an epoxy-based polymer, SU-8, with 50-µm-thick walls. Arrays of 25-nm wide slots were created in the SU-8 microcontainer lid. Isolated mouse islets were encapsulated in the microcontainer, and their physiological response to glucose was studied with fluorescence and two-photon imaging over 48 hours. The physiological response of the encapsulated islets was indistinguishable from controls. An agarose-filled microcontainer was imaged with magnetic resonance imaging to demonstrate the feasibility of future noninvasive, in vivo imaging. The SU-8 microcontainers maintained mechanical integrity upon islet loading and mechanical manipulation. Islet encapsulation, as well as the ability to visualize islet function within these transparent microcontainers, was demonstrated.

Mechanically robust, cell encapsulating microdevices fabricated using photolithographic methods can lead to more efficient immunoisolation in comparison to cell encapsulating hydrogels. There is a need to develop adhesive bonding methods which can seal such microdevices under physiologically friendly conditions. We report the bonding of SU-8 based substrates through (i) magnetic self assembly, (ii) using medical grade photocured adhesive and (iii) moisture and photochemical cured polymerization. Magnetic self-assembly, carried out in biofriendly aqueous buffers, provides weak bonding not suitable for long term applications. Moisture cured bonding of covalently modified SU-8 substrates, based on silanol condensation, resulted in weak and inconsistent bonding. Photocured bonding using a medical grade adhesive and of acrylate modified substrates provided stable bonding. Of the methods evaluated, photo-cured adhesion provided the strongest and most stable adhesion.

We report fabrication and characterization of a magnetically-assembled polymeric nanoporous microcontainer for cell transplantation device. SU-8 cuboid base with 200 x 200 x 200 µm 3 cell loading space was fabricated. In addition, SU-8 lid was fabricated in a manner that it has triple layer with 350 nm thick membrane where myriads of 7~20 nm wide nanoslots were defined by nanoimprint lithography. In both SU-8 base and the lid, another layer of SU-8 with electroplated CoNiMnP permanent magnets with aligned magnetization were fabricated and they were bonded. Pancreatic islet cell was loaded into the SU-8 cuboid base in sterile condition and the lid was assembled by magnetic force. The magnetically-assembled polymeric cell transplantation device was successfully tested with FM -64 dye (~1 nm) for semi-permeability which is essential to the immuneisolation characteristics.