OMENTAL TRANSPLANTATION and CELL CULTURE. by: ROSENDO CRIOLLOS, M.D. A thesis submitted to the faculty of Graduate Studies and Research in partial fulfillment of the requirements of the Master of Science Degree. Department of Experimental Surgery, McGill University, MONTREAL. APRIL 1964. (i) P R E F A C E. The tremendous progress in medicine, especially in cardiovascular surgery during the pBst few decades bas promoted development of measures for the control and cure of various anomalies and diseases by surgical means. While the controversy over the different procedures of revascularization for an ischaemic heart still continues, the rate of surgery in the management of the occlusive coronary artery disease is widely accepted; as James Bryce so ably said, WMedicine is the only profession that labours incessantly to destroy the reason for its own existence". If this be true allow me to thank Dr. Arthur Vineberg for letting me be one of the labourera in his investigations on free omental grafts as a method to revascularize the ischaemic heart. For the 1~ years that I have expended in the field of research under his supervision I wish to extend my appreciation of his thought provoking discussions on the problems encountered throughout this investigation that lead me to develop a method of scientific thinking. These studies afforded me the opportunity to explore more fully the vascularization activities of the omental tissue in re-establishing itself while set free in ectopie environments which resulted in the finding of a (ii) three day minimum of such free omental grafts to become revascularized. The work certainly has roused my interest and enthusiasm in the importance of experimental medicine, enabling me to complete the training in general surgery with better perspective and understanding. It has been a pleasure and enjoyment to do this work in the fascinating fields of cardiac revascularization, tissue transplantation and tissue culture. Here I must express my gratitude to Dr. D. R. Webster, Chairman of the Department of Experimental Surgery, for permitting this work to be carried out in the laboratories, his interest in cardiac revascularization and kindly spirit. To the Director of Research, Dr. S. c. Skoryna, for his gracious being and willingness to help has always been a source of inspiration. The candidate wishes to thank Dr. A. M. Masson, Associate Professer of the Department of Bacteriology, McGill University for her enthusiastic suggestions and guidance on the tissue culture techniques. Special acknowledgement is due to Dr. W. J. Pirozynski, Associate Professer of Pathology, for his interpretations and criticism of the slides as well as the unflogging zeal with which he supported the pursuit of this work. (iii) I am indebted to Dr. S. M. Ban~ill, Pro~essor of Anatomy, for the opportunity of relearning my anatomy while a demonstrator to the first year medical students at the University. I am obliged to Mr. Nagy and his staf~ for their assistance in the operating rooms and care of the dogs. To Mr. Farrell and Mr. s. Byers ~or the eare o~ the small animals. To Mr. C. Hodge and Mr. P. Morin for their photographie work and Mr. J. Szarmany ~or radiologieal aid. I am grateful to Miss N. Johnston, librarian at the Medical Sehool, for her valuable help in the review of the literature and supply o~ copies from libraries abroad and to Miss Nicole Pineau in the preparation of the microscopie sections. I wish to thank Miss B. Murrell for her devoted time and effort in the preparation of this manuscript. Miss c. Fraser deserves a special thanks for her assistance am the typing of my seminar. Finally the candidate wishes to extend his sincere gratitude ~or the interest, ideas and support rendered to me by all members and colleagues of the Department, especially to Dr. D. W. Edwards whose eriticism and talent was weleome and Dr. T. M. Paul whose adviee and ideas were most helpful. (iv) I owe much to Dr. Alan A. Kane of Brooklyn, New York, it would be incomplete without expressing my indebtedness for the opportunities and encouragement given to me during the course of my surgical residency training under his supervision. R. CRIOLLOS, M.D. -: TABLE OF CONTENTS .- Page No. PREFACE ................................................... (i)-(iv) CHAPTER I: INTRODUCTION and PURPOSE •••••••••••••••••••• 1. CHAPTER II: REVIEW OF LITERATURE • • • • • • • • • • • • • • • • • • • • • • • • 6. CHAPTER III: EXPERIMENTAL STUDIES and RESULTS ••••••••••• 16. EXPERIMENT I: Free Omental Transplants in body cavities and subcutaneous tissues •••••••••••••••••••••••• 17. Series 1: Intraperitoneal free omental grafts • • • • • • • • • • • • • • • • • • • • • • 18. Series 2: Intrapleural free omental grafts. 19. Control studies of transplants with other tissues basides omentum ••••••••••••••••••••• 20. Series 3: Intraperitoneal free grafts of omentum, muscle and fat ••••• 22. Series 4: Intrapleural free grafts of omentum, muscle and fat ••••• 24. Series 2: Subcutaneous free omental grafts 26. Series 6: Intrapericardial free omental grafts •••••••••••••••••••••• 28. EXPERIMENT II: Free Omental Transplant in Diffusion Chambers ••••••••••••• 33. Part 1 : In the Transparent Chamber in the dorsal skin of the mouse. 34. Part 2 : In the Anterior Chamber of the Rabbit's Eye •••••••••••••••• 47. EXH:RIMENT III: Omental Cell Culture •••••••••• 57. 1. In Natural Media. 2o In Artificial Media. 3. Cell Suspension Method. Results •••••••••••••••••••••••• 64 (Table of Contents- contd.) Page No. EXFERIMENT IV: Phagocytic Properties of Free Omental Grafts ànd Mast Cella ••••••••••••••••••••• 70. EXFERIMENT V: Cardiac Revascularization with Free Qnental Grafts and Chemical Epicardiectomy •••••••• 73. Methods • • • • • • • • • • • • • • • • • • • • • • • • • • • 75. Control Series •••••••••••••••••••• Resulta • • • • • • • • • • • • • • • • • • • • • • • • • • • 82. CHAPTER IV: C<J.fivlENTS • • • • • •• •• •• •• • • . • • • • ••• • • • ••• • •• •• ••• •• • • 85. CHAPTER V: SU~~RY •••••••••••••••••••••••·••••••••••••·••••• 88. CHAPTER VI: CONCLUSIONS ••••••••••·••••••••••••••••••••••••••• 96. CHAPTER ~ BIBLIOGRAPHY ••••••••••••••••••••••••••••••••••••• 98. -: TABLE OF FIGURES :- Fig. Page No. No. 1. A Free intraperitoneal omental graft, 25 days old • • • • 19 2. Same omental graft with its attachments •••••••••••••• 19 3. A free intrapleural omental graft, 25 days old ••••••• 21 4. A free intrapleural omental graft with its attachments . • . • . • . • • . • 21 5. Photomicrograph of Rat No. 24 with 25 days of intrapleural free omental graft ••••••••••••••••• 23 6. Photomicrograph of section of intrapleural free omental graft •••••••••••••••••.••••••.•••..••.•• 23 7. Photomicrograph of section of intrapleural free omental gra ft . 2 5 8. Rat with intraperitoneal 3 grafts, omentum, muscle and fat . • . • . • . • • • • 25 9. Rat with intrapleural 3 grafts, omentum, muscle and fat . • . 27 10. Subcutaneous free omental graft, 4 weeks old • •••••••• 27 11. Subcutaneous free omental graft, 6 weeks old • • • • • • • • • 29 12. 5 Free omental grafts inside pericardium of dog •••••• 29 13. Same grafts after injection with Schlesinger Mass • • • • 31 14. Section of sorne grafts showing the mass and blood cells in a large omental vessel ••••••••••••••••• 31 15. Intrapericardial free omental grafts attached to internal mammary vessels and heart •••••••••••••• 32 16. Cham ber parts • . • . • . • • . • 3 5 17. Diagram of a cross-section of chamber and graft •••••• 35 18. Mouse with assembled chamber, front view ••••••••••••• 37 19. Mouse with assembled chamber, back view •••••••••••••• 37 (Table of Figures - contd.) Fig. Page No. No. 20. Diagram of 18 chambers in mice .. .. .. 40 21. Chamber with 3 omental transplants, 5 days old •••••• 43 22. Same chamber, same grafts, 10 days old •••••••••••••• 43 23. Photomicrograph of a section of omental graft in the chamber . • • . • . • . 45 24. Diagram of technique of intraocular transplantation.. 45 25. Intraocular omental graft, 3 days old ••••••••••••••• 52 26. Intraocular omental graft after author's injection •• 52 27. Photomicrograph of section of injected intraocular graft • . • . 53 28. Close-up of same graft to see dye in omental vessels. 53 29. Photomicrograph of blood supply of iris after injection . 55 JO. Photomicrograph of blood supply of iris and graft.... 55 31. Contents of Medium No. 150. ••••••••••••••••••••••••• 65 32. Leighton Tube with omenta1 exp1ants • • • • • • • • • • • • • • • • • 66 JJ. Fibro-b1ast 1ike ce11s growing from explants, 2nd day of incubation •••••••••••••••••••••••••• 66 34. Same exp1ants after 10 days of incubation • • • • • • • • • • • 68 35. Tubes with exp1ants at 3, 6 and 9 days of incubation •.........•............•...........•. 68 36. Cat's omentum after intraperitonea1 injection of carbon suspension •••••••••••••••••••••••••••••• 71 37. Photomicrograph of intraperitonea1 free omenta1 graft treated with intraperitonea1 injection of carbon suspension ••••••••••••••••••••••••• ·•• 71 38. Mast cel1s in omenta1 tissue •••••••••••••••••••••••• 74 39. Diagram of free omenta1 graft operation ••••••••••••• 74 (Table of Figures- contd.} Fig. Page No. No. 40. Ameroid constrictor on branches of left coronary artery • • . • 77 41. Ameroid constrictor on right coronary artery •••••••• 77 42. Chemical epicardiectomy .. .. .. .. .. .. .. 78 43. Chemical epicardiectomy • •••••••••••••••••••••••••••• 78 44. Heart after being epicardiectomized ••••••••••••••••• 79 45. Root of aorta after cleaning ••••••••••••.•••••••••••