Direct cell-cell interaction of cardiomyocytes is key for bone marrow stromal cells to go into cardiac lineage in vitro Shinya Fukuhara, MD, Shinji Tomita,

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Direct cell-cell interaction of cardiomyocytes is key for bone marrow stromal cells to go into cardiac lineage in vitro Shinya Fukuhara, MD, Shinji Tomita, MD, PhD, Seiji Yamashiro, MD, MSc, Takayuki Morisaki, MD, PhD, Chikao Yutani, MD, PhD, Soichiro Kitamura, MD, PhD, Takeshi Nakatani, MD, PhD The Journal of Thoracic and Cardiovascular Surgery Volume 125, Issue 6, Pages 1470-1479 (June 2003) DOI: 10.1016/S0022-5223(02)73610-6

Figure 1 Experimental culture system with rat cardiomyocytes (CM) and GFP-BMCs. The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 2 The morphology of GFP-BMCs in passage 2 (A and B) and rat cardiomyocytes (CM; C and D) in vitro. A, These cells were spindle, oval, wedge, or sheet shaped. B, All cells expressed green under fluorescent microscopy. (Original magnification 200×.) D, None of the cells were visible under fluorescent microscopy. (Original magnification 200×.) The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 3 GFP-BMCs with rat cardiomyocytes (CM) on day 2 after coculture in group 3. GFP-BMCs were spindle shaped, attached to cardiomyocytes, and contracted synchronously with cardiomyocytes. (Original magnification 200×.) The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 4 The proliferation of GFP-BMCs in groups 1 (solid line) and 3 (dotted line) is shown. There was no difference between groups 1 and 3 (NS). The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 5 Myogenic differentiation of GFP-BMCs. Cells were stained with a first antibody against MHC. A was photographed during the contrast phase. B and C were photographed during the fluorescent phase (B, green, excitation at 515–540 nm; C, red, excitation at 574–640 nm). D was the double-labeled cell superposition of B and C. Positive cells showed a cross-striated pattern. Combined green and red fluorescence represented myogenic cells derived from BMCs. (Original magnification 400×.) The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 6 The percentage of MHC-positive BMCs from day 0 through day 5 in group 3. The bar represents mean and SE. MHC-positive GFP-BMCs appeared from day 1. There was a significant difference among days in group 3 (P < .0001). As the days passed, the expression of MHC-slow significantly increased to 2.5% on day 5. The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 7 ANP-positive BMCs. Cells were stained with a first antibody against ANP. The phases (A–D) were the same microscopic conditions seen in Figure 5. (Original magnification 400×.) The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 8 The percentage of ANP-positive BMCs in group 3. The bar represents mean and SE. Positive cells appeared on days 2, 3, and 4. Among days, the difference in the percentages were recognized as significant (P = .039). The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 9 Connexin 43-positive BMCs. Cells were stained with a first antibody against connexin 43. Phases A and B were the same microscopic conditions as in Figure 5. Connexin 43 was detected at the margin of BMCs in phase C. (Original magnification 600×.) The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 10 TnI-positive BMCs. Cells were stained with a first antibody against TnI. The phases (A–D) were the same microscopic conditions as in Figure 5. Some BMCs stained positively and showed myofibrils lengthwise along the cell. The expression of TnI corresponded to green fluorescence derived from BMCs. (Original magnification 400×.) The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

Figure 11 The percentage of cardiac-specific TnI-positive BMCs in group 3. The bar represents mean and SE. Positive cells appeared on day 4 and increased to 1.9% on day 5. The differences of the positive BMCs were recognized to be significant among days (P < .0001). The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)

The Journal of Thoracic and Cardiovascular Surgery 2003 125, 1470-1479DOI: (10.1016/S0022-5223(02)73610-6)