Mark F. Fillinger, MD, Susan E. O'Connor, MD, Robert J

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The effect of endothelial cell coculture on smooth muscle cell proliferation Mark F. Fillinger, MD, Susan E. O'Connor, MD, Robert J. Wagner, BS, Jack L. Cronenwett, MD Journal of Vascular Surgery Volume 17, Issue 6, Pages 1058-1068 (June 1993) DOI: 10.1016/0741-5214(93)90676-D Copyright © 1993 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 A, Diagram of Falcon coculture insert with 13 μm thick, porous, PET membrane. B, Schematic diagram of coculture system illustrates EC/SMC experimental group. C, Cross section of EC/SMC coculture (light microscopy; original magnification ×500). Confluent EC are on lower surface, separated from test SMC by 13 μm thick membrane (M). Journal of Vascular Surgery 1993 17, 1058-1068DOI: (10.1016/0741-5214(93)90676-D) Copyright © 1993 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 SMC proliferation rate indexed by thymidine incorporation adjusted for total cellular DNA. Days 1 to 4 indicate time after release from growth arrest in serum-free media. Experimental group (EC/SMC coculture with confluent EC opposite membrane from SMC) is compared with controls of SMC opposite SMC (SMC/SMC) and SMC alone (Ø/SMC). Journal of Vascular Surgery 1993 17, 1058-1068DOI: (10.1016/0741-5214(93)90676-D) Copyright © 1993 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 SMC density indexed by SMC DNA. Days 1 to 4 indicate time after release from growth arrest in serum-free media. Experimental group (EC/SMC coculture with confluent EC opposite membrane from SMC) is compared with controls of SMC opposite SMC (SMC/SMC) and SMC alone (Ø/SMC). Journal of Vascular Surgery 1993 17, 1058-1068DOI: (10.1016/0741-5214(93)90676-D) Copyright © 1993 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Surface view of (A) EC/SMC, (B) SMC/SMC, and (C) Ø/SMC cultures with light microscopy demonstrates differences between test SMC in different cultures. Note thin spindle shape of SMC in EC/SMC group. (Toluidine blue; original magnification × 250.) Journal of Vascular Surgery 1993 17, 1058-1068DOI: (10.1016/0741-5214(93)90676-D) Copyright © 1993 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 5 Electron micrographs of (A) EC/SMC, (B) SMC/SMC, and (C) Ø/SMC cultures in cross section. Cells are separated by porous membrane (M), 13 μm thick with 0.45 μm pores. Pores (P) do not seem to penetrate completely across membrane because sections are not exactly perpendicular to pores. (Original magnification × 2000.) Journal of Vascular Surgery 1993 17, 1058-1068DOI: (10.1016/0741-5214(93)90676-D) Copyright © 1993 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 6 A, Electron micrograph demonstrates cytoplasmic projection (CP) from SMC penetrating membrane pore (P) that appears to extend across entire membrane thickness. (Original magnification × 4500.) B, Higher magnification of different SMC cytoplasmic projection demonstrates cellular elements within projection. (Original magnification × 15000.) Journal of Vascular Surgery 1993 17, 1058-1068DOI: (10.1016/0741-5214(93)90676-D) Copyright © 1993 Society for Vascular Surgery and International Society for Cardiovascular Surgery, North American Chapter Terms and Conditions