Establishment of model human stem cell of transplantation into myocardium of chick embryos

OBJECTIVE TO ESTABLISH AN ANIMAL MODEL OF HUMAN STEM CELL TRANSPLANTATION INTO MYOCARDIUM IN CHICK EMBRYOS. METHODS HOECHSST 33 258 LABELED HUMAN PRIMORDIAL GERM CELLS (HPGC) WERE MICROSURGICALLY INJECTED INTO THE MYOCARDIUM OF 633 CHICK EMBRYOS OF 3~4 DAYS DEVELOPMENT. TEN DAYS AFTER, THE HEARTS WERE ISOLATED FROM THE 94 SURVIVING CHICK EMBRYOS, EMBEDDED, AND SLICED. IN SITU HYBRIDIZATION (ISH) WITH HUMAN SPECIFIC DNA ALU PROBE WAS CONDUCTED ON THE SECTIONS WITH FLUORESCENCE TO DETECT THE EXISTENCE OF TRANSPLANTED PGC. IMMUNOHISTOCHEMISTRY WITH HUMAN-MYOCARDIUM-SPECIFIC ANTIBODY CTNT WAS CONDUCTED ON THE ADJACENT SECTIONS TO OBSERVE THE DIFFERENTIATION OF HUMAN MYOCARDIAL CELLS. RESULTS ISH SHOWED THAT PGC WERE DETECTED IN THE MYOCARDIUM OF CHICK EMBRYOS 10 DAYS POST-OPERATIONALLY. IMMUNOHISTOCHEMISTRY SHOWED THAT THE MYOCARDIUM ADDED WITH ANTIBODY IN ADJACENT SECTIONS WAS CTNT-POSITIVE AND THE MYOCARDIUM UNTREATED WITH ANTIBODY WAS CTNT-NEGATIVE. SUCCESSFUL CELL TRANSPLANTATION OCCURRED IN 15.3%±2.4% OF CHICK EMBRYOS. CONCLUSION ESTABLISHMENT OF AN ANIMAL MODEL OF CELL TRANSPLANTATION OF HUMAN STEM CELLS INTO MYOCARDIUM IN CHICK EMBRYOS IS FEASIBLE.