Using imbibition of macroporous polyacrylamide with aminoacids, as well as combined system of alginate and collagen, we, together with our colleagues from the Institute of Bioorganic Chemistry and the Institute of Physics, have developed a new type of cells carrier (Patent BY 8699 C1). This allowed proper immobilization and proliferation of cells without a significant decrease in their viability, which was previously observed mainly due to cell clamping. We align cells using gradient electromagnetic fields (see in detail) and then tested viability of cells in 2 weeks after the immobilization. The cells viability was found to be preserved, and the development of the satellite fibroblast-like elements was negligible. A reliable increase (31.6 ± 9.7%) in amount of hepatocytes was observed, their viability being 97-98%. There were neither annexin-positive no CD-95-positive cells.

To analyze the possibility of development and cooperation of cells inside the implant, we tested implantation plates made of materials, which offer both cells protection and support. We used acrylamide and alginate to coat the transplanted cells, as these materials were found to be bio-inert in experiments with animals. To form the inner layer, which contains growing cells, collagen and 7% of agarose were tested. We also took into account adhesion possibilities and marginal thickness of material to provide adequate trophic conditions. Besides, we tested new modifications of acrylamide to produce anti-fibroblast and immuno-isolating coating of plate. Polyamide was used to produce the frame of implantation plate.

More than 30 fragments of implantation plates have been investigated. We performed 107 tests of cell viability using Annexin kit. As much as 16 cultures of cells from fetal porcine pancreas and liver were studied. The results obtained suggest that cells inside laminar containers, which were preliminarily affected with different physicochemical factors to stabilize alignment of cells, preserve high level of viability.

Conclusions.

• A сompletely new type of cells carrier has been developed using imbibition of macroporous polyacrylamide with aminoacids. This allowed proper immobilization and proliferation of cells without a significant decrease in their viability. The number of functional elements (islet cell clusters) per square unit has increased to 240 ± 48%.
• The investigation of implantation plates with polyacrylamide coating showed apparent anti-fibroblast withstandability of the plate made according to Trewan and Grower, 1997. The plate overgrown with fibroblasts was less than 15% of total surface.
• Additional immobilization of PA-peptide in acrylamide stitch leads to considerable increase in viability and provides adequate functional activity of cells in the organism of recipient animal.
• Acrylamide plate of thickness more than 75 μm protects β-cells from the damage by C3-component of complement in the serum obtained from the patients with IDDM, protection from cytokines being not detected.
• Surgical methods of implantation of containers with hepatocytes and insulocytes to the laboratory animals have been worked out. Complete compensation of alloxan-induces diabetes was archieved.
• The efficiency of additional coating of container with immunoprotective and anti-fibroblast materials has been studied. When transplanting into perivascular and subcapsular sites this additional coating was showed to be quite effective.

Journal of Clinical Investigation (in press)