Protocol for Transplantation of Healthy Cells Between Adult Drosophila of Different Ages and Sexes

Abstract

Drosophila melanogaster serves as a powerful and versatile model organism for studying tissue allotransplantation due to its short life cycle, genetic manipulability, and significant homology to mammalian signaling pathways. This protocol outlines a procedure for performing tissue transplants between adult individuals of different ages and sexes. Key steps include dissection of the donor’s midgut tissue, microinjection into the recipient, and tracking engraftment using sex chromosome differences. The protocol demonstrates high short-term survival (over 80%) of host organisms, with transplanted tissues encapsulated by hemocytes. Sexual dimorphism affects transplant outcomes, with females showing stronger immune responses through the Toll pathway, resulting in more frequent rejections, while males exhibit greater tissue tolerance. Age-related factors, including reduced regenerative capacity and oxidative stress in older individuals, impact transplantation success. This methodology also enables modeling of intestinal stem cell regeneration. Despite challenges such as small tissue size and lack of an adaptive immune system, the protocol offers valuable insights into innate immunity, aging, and intercellular interactions, positioning Drosophila as an ideal preclinical model for studying tissue regeneration and immune response.