Transplantation of Murine Bone Marrow Stromal Cells under the Kidney Capsule to Secrete Coagulation Factor VIII

Abstract

Ectopic cell transplantation has been studied as an alternative to whole organ transplantation or as a method to produce secretable proteins for genetic disorders. In this study, bone marrow stromal cells isolated from C57Bl/6 mice were genetically modified to express either lacZ-or B-domain-deleted human factor VIII. In vitro modification of the isolated bone marrow stromal cells was initially performed by transducing increased doses of VSV-G pseudotyped lentiviral vectors expressing lacZ. At a MOI of 25, all of the bone marrow stromal cells were X-gal positive, which maintained their ability to expand and differentiate prior to transplantation into mice. Extremely poor engraftment was observed in the liver, but transplantation of the bone marrow stromal cells expressing lacZ under the kidney capsule resulted in long-term viable X-gal-positive cells for at least 8 weeks (length of study). In vitro expression of human factor VIII was detected in a dose-dependent manner following bone marrow stromal cell with a factor VIII-expressing lentiviral vector. Transplantation of the factor VIII-expressing bone marrow stromal cells under the kidney capsule led to long-term therapeutic expression in the mouse plasma (1-3 ng/ml; n = 4-5 mice/group) for 8 weeks. This study demonstrated that ectopic transplantation of bone marrow stromal cells under the kidney capsule can be effective as a method to express secretable proteins in vivo.

Genetically Modified Adipose Tissue-Derived Stem/Stromal Cells, Using Simian Immunodeficiency Virus-Based Lentiviral Vectors, in the Treatment of Hemophilia B

“…For this reason, genetic methods are required to safely transport genes of interest into the cells to mass produce intracellular or secreted proteins or to inhibit the transcription/translation of deleterious genes. Earlier studies have shown some promise in the use of genetically modified bone marrow-derived mesenchymal stem cells (BM-MSCs) as a possible cell-based treatment modality for hemophilia (Oh et al, 2006;Coutu et al, 2011). Compared with BMMSCs, ADSCs may have an advantage in that they are abundant and can be obtained by less invasive procedures (Kern et al, 2006).…”

“…complex retroviruses (Coffin et al, 1997;Walther and Stein, 2000;Anjos-Afonso et al, 2004;Haleem-Smith et al, 2005;Oh et al, 2006;Talens-Visconti et al, 2006;Zaragosi et al, 2007;Li and Lu, 2009;Sugii et al, 2010;Coutu et al, 2011;Kim et al, 2011;Li et al, 2011b).…”

“…Cell-based therapies have received a great deal of attention as a next-generation therapeutic approach for hemophilia (Oh et al, 2006;Follenzi et al, 2008;Kasuda et al, 2008;Tatsumi et al, 2008a,b;Ohashi et al, 2010). There has been enormous interest in the transplantation of stem cells to produce clotting factors (Chuah et al, 2004;Oh et al, 2006;Coutu et al, 2011).…”

“…There has been enormous interest in the transplantation of stem cells to produce clotting factors (Chuah et al, 2004;Oh et al, 2006;Coutu et al, 2011). Some types of stem cells can be readily isolated from human patients with minimal invasiveness (Lin et al, 2008), such as adipose tissue-derived stem/stromal cells (ADSCs, also known as adipose tissue-derived mesenchymal stem cells, AT-MSCs) (Zuk et al, 2001;Li et al, 2011b).…”

“…A multitude of genetic approaches (Anjos-Afonso et al, 2004;Haleem-Smith et al, 2005;Oh et al, 2006;Talens-Visconti et al, 2006;Sugii et al, 2010;Coutu et al, 2011;Li et al, 2011a,b) have been applied to exogenously produce clotting factors.…”