JP-7856990-B2 - Evaluation methods for transplant materials

Inventors

• 宮内 浩

Assignees

• 宮内 浩
• 宮内 久美子
• 宮内 真弥子
• 宮内 琉美子
• 宮内 悠慧

Dates

Publication Date

20260512

Application Date

20250120

Claims (10)

1. A step of obtaining a recipient immune to the donor by subcutaneously administering a crude protein solution extracted from the cells or tissues of a labeled marker-expressing donor to a newborn recipient, An immune-tolerant recipient (excluding humans) produced by the step of implanting a transplant material containing an artificial transplant material in which tissue or cells of a donor expressing the labeling marker are covered, adhered to, infiltrated, or incorporated into the immune-tolerant recipient, wherein the donor and the recipient are inbred animals, and the method of producing an immune-tolerant recipient and observing the behavior of the labeling marker in the immune-tolerant recipient, A method comprising, thereby identifying whether cells and tissues generated in the immune-tolerant recipient after transplantation originate from the immune-tolerant recipient or from the donor.
2. A step of obtaining a recipient immune to the donor by subcutaneously administering a crude protein solution extracted from the cells or tissues of a labeled marker-expressing donor to a newborn recipient, An immune-tolerant recipient (excluding humans) produced by the step of transplanting a transplant material containing an artificial transplant material in which tissue or cells of a donor expressing the labeling marker are covered, adhered to, infiltrated, or incorporated into the immune-tolerant recipient, wherein the donor and the recipient are inbred animals, and the behavior of the labeling marker in the immune-tolerant recipient, as well as the response and progress of the transplant model animal, A method comprising, thereby confirming the state of immune tolerance in a recipient of the transplanted material after transplantation.
3. A method for identifying whether cells and tissues generated in an immune-tolerant recipient after transplantation are derived from the immune-tolerant recipient or from the donor, according to claim 1, wherein the transplant material is a tubular structure, a membranous structure, or an artificial organ.
4. A method for identifying whether cells and tissues generated in an immune-tolerant recipient after transplantation are derived from the immune-tolerant recipient or the donor, as described in claim 1, wherein the transplant material is retrieved after transplanting the transplant material into the donor's body and leaving it in place for 2 to 30 days.
5. A method for identifying whether cells and tissues generated in an immune-tolerant recipient after transplantation are derived from the immune-tolerant recipient or from the donor, according to claim 1, wherein the labeling marker is a fluorescent protein.
6. The method for identifying whether cells and tissues generated in an immune-tolerant recipient after transplantation are derived from the immune-tolerant recipient or from the donor, wherein the transplant material is a collagen sheet on which donor-derived cells have been seeded.
7. A method for confirming the state of immune tolerance in a recipient of a transplanted material according to claim 2, wherein the transplanted material is a tubular structure, a membranous structure, or an artificial organ.
8. A method for confirming the state of immune tolerance in a recipient of a transplanted material according to claim 2, wherein the transplanted material is obtained by transplanting the transplanted material into the living body of the donor and leaving it in place for 2 to 30 days before retrieval.
9. A method for confirming the state of immune tolerance in a transplant recipient after transplantation of a transplant material, according to claim 2, wherein the labeling marker is a fluorescent protein.
10. The method for confirming the state of immune tolerance in a recipient of a transplanted material according to claim 2, wherein the transplanted material is a collagen sheet on which donor-derived cells have been seeded.

Description

This invention relates to a method for evaluating transplant materials that can be used as a highly reliable and reproducible approximate autologous transplant experimental system and that exhibit high homology to clinical medicine. Regenerative medicine is a technology that repairs or replenishes the function and/or structure of living tissues in humans, livestock, and pets when that function and/or structure is damaged or missing. Furthermore, regenerative medicine is also useful in cases where growth is inhibited due to disease or injury during the developmental or growth period of animals, resulting in the failure to achieve the desired function and/or structure. One of the most effective methods in regenerative medicine involves transplanting bio-derived medical materials, including cells or tissues, into the deficient area. In Japan, such medical materials are defined as "regenerative medicine products" under the three laws related to regenerative medicine (the Regenerative Medicine Promotion Act, the Regenerative Medicine Safety Assurance Act, and the amended Pharmaceutical Affairs Act). From an ethical and immunological standpoint, regenerative medicine products used in transplantation should preferably be manufactured using materials derived from the recipient individual, a practice known as "autoplastic transplantation." Furthermore, artificial materials are used to complement elements that are insufficient in medical materials derived from living organisms alone. Such artificial materials must not cause undesirable reactions in the body, including immune responses. Conventionally, various medical materials have been used, including not only bio-derived materials such as regenerative medicine products, but also biocompatible materials and bio-compatible materials (for example, Patent Documents 1-4). However, it is difficult to verify whether these materials are optimal in terms of material, physical properties, and composition before transplantation. In particular, it has been difficult to verify the performance of regenerative medicine products containing living cells and tissues in advance. To ensure the quality of regenerative medicine products, a clinically relevant system is necessary that allows for prior animal testing using mammals and monitoring the progression of immune responses after transplantation. For such animal testing, an animal model employing "autologous transplantation," where cells and tissues are collected from the organism intended for transplantation and returned to that organism, is suitable. However, animal experimental systems using autologous transplantation have been rarely used as a research tool until now. In particular, autologous transplantation models for small animals have practically nonexistent. This is because autologous transplantation is difficult in small animals, including mice and rats. Even if the amount of cells or tissue collected for transplantation is less than 1 gram, small animals may not be able to withstand the collection procedure, or depending on the tissue, the amount may be lethal. In small animals, tissues and cells are generally collected after the animal is killed and bled. Therefore, the animal is lost during the collection procedure, making autologous transplantation impossible. While autotransplantation is possible with medium and large animals, the high cost of raising them makes it difficult to conduct sufficient experiments. Furthermore, with the growing awareness of animal welfare, there is a trend towards stricter regulations on animal experimentation itself. Therefore, much of the conventional research has been built around experimental systems centered on allograft and heterograft/xenograft in small animals. Since allograft and xenograft are not autotransplants, the use of immunosuppressants, or the use of severely immunodeficient animals (nude mice, NOD/scid mice, NOG mice, etc.), or a combination of both, is necessary to avoid immune responses. This necessitates the use of individuals that are expensive to breed and maintain, resulting in high costs and labor. Furthermore, these experimental systems require highly specialized facilities, such as sterile rooms, due to the increased risk of infection from immunosuppressant use or immunodeficiency. Additionally, the need for long-term use of immunosuppressants further increases costs. Furthermore, a significant problem with immunodeficiency experimental systems is that experiments are conducted under conditions that are far removed from clinical practice in medicine and veterinary medicine. In particular, in humans, patients with immunodeficiency or those on long-term immunosuppressant therapy are rarely candidates for transplantation. Treatment methods established in immunodeficiency experimental systems may cause unforeseen immune responses when introduced into patients with normal immune function or livestock. Therefore, this invention provides an animal experiment system that