Fibrin glue increases the cell survival and the transduced gene product secretion of the ceiling culture-derived adipocytes transplanted in mice

Abstract

The development of clinically applicable scaffolds is important for the application of cell transplantation in various human diseases. The aims of this study are to evaluate fibrin glue in a novel protein replacement therapy using proliferative adipocytes and to develop a mouse model system to monitor the delivery of the transgene product into the blood and the fate of the transduced cells after transplantation. Proliferative adipocytes from mouse adipose tissue were transduced by a retroviral vector harboring the human lecithin-cholesterol acyltransferase (lcat) gene, and were subcutaneously transplanted into mice combined with fibrin glue. The lcat gene transduction efficiency and the subsequent secretion of the product in mouse adipocytes were enhanced using a protamine concentration of 500 μ g/ml. Adipogenesis induction did not significantly affect the lcat gene-transduced cell survival after transplantation. Immunohistochemistry showed the ectopic enzyme production to persist for 28 days in the subcutaneously transplanted genetransduced adipocytes. The increased viability of transplanted cells with fibrin glue was accompanied with the decrease in apoptotic cell death. The immunodetectable serum LCAT levels in mice implanted with the fibrin glue were comparable with those observed in mice implanted with Matrigel, indicating that the transplanted lcat gene-transduced adipocytes survived and functioned in the transplanted spaces with fibrin glue as well as with Matrigel for 28 days. Thus, this in vivo system using fibrin is expected to serve as a good model to further improve the transplanted cell/scaffold conditions for the stable and durable cell-based replacement of defective proteins in patients with LCAT deficiency.

Effect of Fibrin Formulation on Initial Strength of Tendon Repair and Migration of Bone Marrow Stromal Cells in Vitro

“…They concluded that the most favorable combination for survival of bone marrow stromal cells was 20 mg/mL of fibrinogen and 87 NIH units/mL of thrombin, values intermediate between those of our control and low-low groups. The ratio of dead to total cells in our study was <30%, a finding consistent with previous reports 18,27 . Gugerell et al reported that thrombin concentrations of >400 NIH units/mL induced apoptosis of human keratinocytes 27 .…”

“…The results of these reports have not been consistent, with some studies demonstrating that fibrin scaffolds increased cell viability, proliferation, or differentiation 18,20 and others showing that fibrin may adversely affect these parameters 16,18,20 . An in vitro tendon repair model showed that fibrin gel, acting as a carrier of growth differentiation factor-5 (GDF-5)-treated muscle-derived stem cells, enhanced tendon healing, as evaluated both mechanically and histologically, compared with a collagen gel at both two weeks and four weeks 21 .…”

“…A number of studies have been performed to investigate the effect of fibrin scaffolds seeded with stem cells with and without growth factor augmentation 16,[18][19][20] . The results of these reports have not been consistent, with some studies demonstrating that fibrin scaffolds increased cell viability, proliferation, or differentiation 18,20 and others showing that fibrin may adversely affect these parameters 16,18,20 .…”

The effect of medium selection on adipose-derived stem cell expansion and differentiation: implications for application in regenerative medicine

“…In this study we isolated and cultured ADSCs in seven proliferation media as described by various authors (Katz et al 2005;Yoshimura et al 2006;Bunnell et al 2008;Du et al 2010;Zhou 2011;Aoyagi et al 2011;Frerich et al 2012) and two other proprietary media. Rate of proliferation is an important factor for clinical application, where autologous cells are required for reconstructive surgery.…”

“…Adipose-derived stem cells (ADSCs) harvested from human adipose tissue provide an abundant potential source of multipotent adult stem cells (Rodbell 1964;Zuk et al 2001Zuk et al , 2002Du et al 2005;Katz et al 2005;Yoshimura et al 2006;Zhu et al 2008;Bunnell et al 2008;Tapp et al 2009;Du et al 2010;Maumus et al 2011;Aoyagi et al 2011;Zhou 2011;Sachs et al 2012). These can be harvested by minimally invasive procedures with a potential for diverse applications, such as studies in stem cell biology as well as clinical applications in regenerative medicine.…”