Transplantation of genetically marked cardiac muscle cells

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Presentation transcript:

Transplantation of genetically marked cardiac muscle cells Satoshi Gojo, MDa, Soichiro Kitamura, MDa, Osamu Hatano, PhDb, Akira Takakusu, MDb, Kenji Hashimoto, PhDb, Yumi Kanegae, PhDc, Izumu Saito, MDc The Journal of Thoracic and Cardiovascular Surgery Volume 113, Issue 1, Pages 10-18 (January 1997) DOI: 10.1016/S0022-5223(97)70394-5 Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 1 Structure of Adex1CALacZ vector. In place of E1A and E1B, an expression unit composed of chicken β-actin promoter with cytomegalovirus enhancer (CAG), β-gal gene (LacZ) complementary DNA, and a polyadenylate signal (pA) was introduced to generate Adex1CALacZ expression vector. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 2 X-gal staining of cultured murine fetal cardiac cells. A, Microscopic photograph of culture 1 day after cells had been plated. Approximately 90% to 95% of cultured cells in each field beat spontaneously. Beating rate in these myocytes was not uniform. Clusters, however, did beat synchronously. X-gal staining was performed 1 day after adenovirus infection. B, All cells had assays positive for β-gal showing blue color, at 10 MOI. Bars indicate 100 μm. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 2 X-gal staining of cultured murine fetal cardiac cells. A, Microscopic photograph of culture 1 day after cells had been plated. Approximately 90% to 95% of cultured cells in each field beat spontaneously. Beating rate in these myocytes was not uniform. Clusters, however, did beat synchronously. X-gal staining was performed 1 day after adenovirus infection. B, All cells had assays positive for β-gal showing blue color, at 10 MOI. Bars indicate 100 μm. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 3 X-gal and HE staining of hearts. Histologic examination of host hearts was carried out each week after implantation. A, Staining with X-gal on postoperative day 21. B, Staining with X-gal 12 weeks after implantation. C, Staining with HE on postoperative day 21. Blue cells aligned themselves along layers of host myocardium. Neither fibrous scars nor infiltrations of neutrofils were detectable within any section. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 3 X-gal and HE staining of hearts. Histologic examination of host hearts was carried out each week after implantation. A, Staining with X-gal on postoperative day 21. B, Staining with X-gal 12 weeks after implantation. C, Staining with HE on postoperative day 21. Blue cells aligned themselves along layers of host myocardium. Neither fibrous scars nor infiltrations of neutrofils were detectable within any section. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 3 X-gal and HE staining of hearts. Histologic examination of host hearts was carried out each week after implantation. A, Staining with X-gal on postoperative day 21. B, Staining with X-gal 12 weeks after implantation. C, Staining with HE on postoperative day 21. Blue cells aligned themselves along layers of host myocardium. Neither fibrous scars nor infiltrations of neutrofils were detectable within any section. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 4 TEM analysis from postoperative day 7. Cells with β-gal possess electron-dense deposits in endoplasm. A, Grafted cells had abundant mitochondria, microtubles, and intercalated disks with host and other grafted cells. Bar indicates 2 μm. B, Desmosomes (DES.) and gap junctions (GAP.J) were clearly found in intercalated disks. Bar indicates 500 nm. +, Grafted cardiomyocyte; −, host cardiomyocyte; N, nucleus. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 4 TEM analysis from postoperative day 7. Cells with β-gal possess electron-dense deposits in endoplasm. A, Grafted cells had abundant mitochondria, microtubles, and intercalated disks with host and other grafted cells. Bar indicates 2 μm. B, Desmosomes (DES.) and gap junctions (GAP.J) were clearly found in intercalated disks. Bar indicates 500 nm. +, Grafted cardiomyocyte; −, host cardiomyocyte; N, nucleus. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions

Fig. 5 Electrocardiograms of mice in transplanted and control groups. Electrocardiograms were recorded before mice in grafted group were killed for histologic and TEM examination. Recording speed was 100 mm/min. A, Preoperative electrocardiogram. B, Electrocardiogram at postoperative week 1. C, Electrocardiogram at postoperative week 12. No significant difference in electrocardiographic pattern was found between grafted and control groups. In addition, no arrhythmia was recorded in grafted group at any time. The Journal of Thoracic and Cardiovascular Surgery 1997 113, 10-18DOI: (10.1016/S0022-5223(97)70394-5) Copyright © 1997 Mosby, Inc. Terms and Conditions