Volume 62, Issue 4, Pages (October 2002)

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Volume 62, Issue 4, Pages 1395-1401 (October 2002) Delivery of erythropoietin by encapsulated myoblasts in a genetic model of severe anemia Christopher Rinsch, Phillipe Dupraz, Bernard L. Schneider, Nicole Déglon, Patrick H. Maxwell, Peter J. Ratcliffe, Patrick Aebischer Kidney International Volume 62, Issue 4, Pages 1395-1401 (October 2002) DOI: 10.1111/j.1523-1755.2002.kid574.x Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 1 (A) Transgenic construct employed for generating the 134.3 LC, Epo-TAgH mouse line. Epo in the untranslated region is in the gray bars and in the translated region is solid black; the large T antigen is represented by the hatched area; Ex is exon. (B) Western blot of the immunoprecipitated SV40 large T antigen and Epo expressed in kidneys of anemic homozygous Epo-TAgH mice (-/-) and wild-type (+/+) littermates. Kidney protein extracts were immunoprecipitated either with antibodies against the SV40 large T antigen or Epo. Cell lysates of bTC-tet cells expressing the SV40 large T antigen were employed as a positive control (hc is heavy chain)10. (C) rtPCR of mouse Epo expressed in the kidney tissue of homozygous (-/-) and heterozygous (+/-) Epo-TAgH mice. The band migrating near 600bp corresponds to the size of the mouse Epo cDNA. Reactions carried out in the absence of primers served as negative controls: (+/-) control and (-/-) control lanes. Kidney International 2002 62, 1395-1401DOI: (10.1111/j.1523-1755.2002.kid574.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 2 Comparison of wild-type (+/+; ○) and anemic homozygous (-/-; •) Epo-TAgH mice to subcutaneously implanted capsules secreting mouse Epo (N = 7). Data are presented as mean ± SEM (*P < 0.05; **P < 0.01). Kidney International 2002 62, 1395-1401DOI: (10.1111/j.1523-1755.2002.kid574.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 3 Two of the homozygous mice maintaining elevated hematocrits were followed for a period of 9 and 12 months prior to explantation (arrows), at which point they returned to their normal baseline hematocrit levels. Symbols are: (•) mEPO; (○) control. Kidney International 2002 62, 1395-1401DOI: (10.1111/j.1523-1755.2002.kid574.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 4 Homozygous anemic Epo-TAgH mice implanted with capsules secreting mouse Epo were treated with anti-CD4+ mAb. Efficacy of (A) transient (days -3, 0, +3) or (B) intermittent dosing (days -3, 0, +3, +14, +28, +42) was compared. Kidney International 2002 62, 1395-1401DOI: (10.1111/j.1523-1755.2002.kid574.x) Copyright © 2002 International Society of Nephrology Terms and Conditions

Figure 5 Photomicrograph of encapsulated C2C12 mEpo myoblasts following 8 weeks in vivo. Clusters of viable myoblasts were observed within the capsule's interior. Kidney International 2002 62, 1395-1401DOI: (10.1111/j.1523-1755.2002.kid574.x) Copyright © 2002 International Society of Nephrology Terms and Conditions