JP2019218273A - Method for creating insulin-producing pancreatic islet cells - Google Patents

Abstract

To provide newly insulin-producing cells in subcutaneous white adipose tissue by allowing exogenous insulin-producing cells to coexist in subcutaneous white adipose tissue.SOLUTION: The present invention newly creates insulin-producing cells derived from a recipient in subcutaneous white adipose tissue by transplanting exogenous insulin-producing cells into subcutaneous white adipose tissue site of a diabetic recipient, for example, into the subcutaneous white adipose tissue in the groin. By allowing the exogenous insulin-producing cells to coexist in the subcutaneous white adipose tissue in which the vegetative blood vessels are widely distributed, a novel insulin-producing cells is produced near the coexisting site in the subcutaneous white adipose tissue.SELECTED DRAWING: None

Description

  The present invention relates to the creation of insulin-producing cells in in vivo tissues other than pancreatic islets in the pancreas.

  Diabetes is a typical lifestyle-related disease, a complex chronic disease in which carbohydrates, lipids and proteins cannot be properly maintained and used. Diabetes is also characterized by abnormally high blood sugar levels due to insulin production deficiency or insulin dysfunction caused by various genetic and environmental factors. The progression of diabetes causes complications such as retinopathy, nephropathy, and neuropathy, causing stroke and cerebrovascular accidents, and consequently, a significant decrease in the quality of life (QOL) of the patient. And ultimately lead to a fatal prognosis for the patient.

  Recently, the number of diabetic patients is estimated to reach about 350 million worldwide (Non-Patent Document 1). Moreover, the total number of diabetic patients, including those strongly suspected of suffering from diabetes and those in the reserve army of diabetes, becomes a huge number, which is clearly a major problem. In addition, the fact that only a limited number of diabetics are receiving adequate treatment is a major medical and economic problem.

  There are two main forms of diabetes, type 1 diabetes (T1DM) and type 2 diabetes (T2DM). Type 1 diabetes (formerly known as insulin-dependent diabetes mellitus (IDDM)) is characterized by the specific destruction of insulin-producing pancreatic beta cells and depletion of insulin. This type 1 diabetes often affects children and young people. Type 2 diabetes (formerly known as non-insulin-dependent diabetes mellitus (NIDDM)) is characterized by insufficient insulin production due to dysfunction of insulin-producing pancreatic beta cells and muscle and liver cells responding properly to insulin produced by the pancreas It is caused by a relative shortage of insulin when it is not possible. It has been noted that this type 2 diabetes occurs most frequently in adults but is also increasing in young people. Type 2 diabetes accounts for about 90% of the total number of diabetic patients. In particular, the number of type 2 diabetic patients is rapidly increasing due to obesity, poor physical activity, and aging. More notably, many young patients with type 2 diabetes have autoimmune abnormalities of the pancreas, making it difficult to distinguish type 2 diabetes from type 1 diabetes or vice versa. That is.

  As described above, patients with type 1 diabetes (T1DM) undergo complete or almost complete destruction of insulin-producing β-cells in the islets of Langerhans (islets) due to infiltration of immune cells, causing rapid fluctuations in blood glucose levels. Events that cause and ultimately lead to complications related to life and death-related hyperglycemia (hyperglycemia) have led to a loss of insulin production capacity. Therefore, patients with type 1 diabetes must rely on insulin therapy by administering insulin multiple times a day or by externally administering insulin via an insulin pump (Non-Patent Document 2).

  However, even when type 1 diabetics receive advanced treatment with insulin, the complications of maintaining stable and sustained blood glucose levels and causing chronic long-term organ damage caused by hyperglycemia Is difficult to prevent. Insulin therapy also causes excessive insulin administration, thereby increasing the risk of developing acute hypoglycemia with significant alterations in consciousness that can be fatal. Despite these difficulties and dangers, insulin therapy now plays a very important role in rescuing type 1 diabetes from emergencies leading to sudden death. However, insulin therapy is just a coping treatment, not a complete cure for type 1 diabetes, and solves very difficult problems such as the risk of hypoglycemia and the development of long-term complications. (See, for example, Patent Document 1).

  As another method of such insulin therapy for treating type 1 diabetes, there is an islet transplantation method in which insulin-producing β cells of Langerhans islets isolated from the pancreas are directly transplanted into a type 1 diabetic patient. This islet transplantation method is theoretically an excellent method for diabetic patients having islet cell disorders, and is an endocrine supplementation therapy that has been widely tried in clinical practice since the late 1960s. This islet transplantation is now a very promising approach to the treatment of type 1 diabetes. This islet transplantation is a simple method of transplanting simply by instilling islet cells into the portal vein of the liver by instillation, so it is extremely safe and dangerous in terms of surgery compared to surgery to transplant all organs. There is an advantage that the procedure can be performed even for a patient having a high possibility. This islet transplant also has the advantage of eliminating the need for abdominal surgery or vascular anastomosis. In addition to this, this islet transplantation has the advantage that, even if a transplant rejection reaction occurs, there is no need to remove it, and the burden on the patient is extremely low (for example, see Patent Document 1).

  However, this islet transplantation method suffers from the disadvantage that the survival rate of transplanted islets is extremely low due to early transplant rejection. Therefore, even if a patient with type 1 diabetes receives islet cell transplantation from the pancreas of one donor, it cannot become insulin-independent (for example, Non-Patent Documents 3 and 4). It is estimated that the rate of early loss of transplanted islet cells is about 60% of the total number of transplanted islet cells. Therefore, in the current islet transplantation, it is necessary to repeatedly transplant islet cells from the pancreas of two or more humans in order to treat one diabetic patient in a favorable condition (for example, see Non-Patent Literature) 3, 4, 5).

  However, this islet transplantation also carries the risk that the islet graft will form an embolus in the peripheral portal vein and implant in the liver of a diabetic patient, causing ischemic alteration in the corresponding area. This can result in a post-transplant immune response, releasing inflammatory santokines that are harmful to islet cell debris. Β-cells that secrete insulin are damaged when exposed to pro-inflammatory cytokines such as, for example, IFN-γ, TNF-α, and IL-1β, and the engraftment rate is significantly reduced (for example, Non-Patent Document 6).

  The first clinical transplant of islet cells into human patients was performed in 1974 at the University of Minnesota. However, its performance was poor until recently compared to other organ transplants. However, in 2000, the University of Alberta released an extremely high rate of insulin under a clinical islet transplantation protocol called the “Edmonton Protocol” using immunosuppression without any steroids. There was a report that this was done (for example, Non-Patent Document 4). Since then, islet transplantation has been established as a way to be a breakthrough in the treatment of type 1 diabetes.

  As noted above, islet transplantation is becoming increasingly useful as a fundamental treatment for diabetes, but further improvements are required for successful islet transplantation. The reason for this is that current islet transplantation, for example, has a low engraftment rate of islet cells, requires a large infusion of islet cells for such a low engraftment rate, and has a small number of donors. Because they still face a number of problems. In 2006, Shapiro et al. (AM Shapiro et al.) Reported that the results of patients with islet transplants using the Edmonton protocol showed that 76% of patients who once did not need insulin therapy after islet transplantation Two years after the transplant, insulin therapy had resumed.

  Furthermore, a major problem is that current islet transplants require more than one donor for a single diabetic recipient due to the low engraftment rate of islet cells transplanted into the liver. This should be resolved urgently, even from the standpoint of scarce donors. If this problem were solved and the ratio of donors to recipients improved to a one-to-one ratio, pancreatic islet transplantation was expected to spread explosively and make a significant contribution to the treatment of diabetes. You.

  Thus, although islet transplantation faces several challenges, decades of research on improving intrahepatic islet delivery have shown that intrahepatic space is not It was found to be a support and to play an essential role in maintaining the engraftment and function of the transplanted islet cells for a long period of time. Therefore, islet transplantation is an attractive method for treating diabetes, and at present, the liver via the portal vein of diabetic patients is a transplant site in a clinical setting (see, for example, Non-Patent Documents 4 and 7). However, on the other hand, there are reports that the hepatic vascular system is a hostile environment that limits engraftment and function (Non-Patent Documents 8 and 9). As a result, a number of alternative sites for islet transplantation were tested to optimize islet engraftment and function, reduce the number of transplanted islet cell clusters needed, and reduce immunogenicity (non- Patent Documents 9 and 10).

  In addition to the intraportal site of the liver, many sites include the subcapsular region of the kidney, spleen, omentum, pancreas, gastrointestinal wall, immunoprotective sites such as the thymus, brain and testis, musculoskeletal sites such as bone marrow, and subcutaneous sites Other transplant sites were examined (eg, Non-Patent Documents 9, 11, and 12). Although many other implantation sites in patients have been tested as described above, to date patients have not been able to be made non-insulin dependent. As a result, intraportal islet transplantation has been empirically recognized as the best site for use in patients (Non-Patent Document 9).

  However, the development of a new transplant site other than the intraportal site of the liver is needed to reduce the loss of islet cells transplanted in diabetic patients and to overcome the problems associated with the shortage of islet cells to be transplanted It is.

  In addition, recent studies have revealed islet cells containing both insulin and glucagon granules. Until now, it was known that insulin is produced in β cells of the islets of Langerhans and glucagon is produced in the α cells (see Non-Patent Document 13). Furthermore, although insulin deficiency was considered to be the only problem in diabetes for decades, it was suggested that glucagon also plays an important role in the pathogenesis of diabetes (Non-Patent Document 14).

  Therefore, these islet cells are expected to play some role in secreting these hormones in vivo once transplanted into diabetic patients.

  On the premise of the background art described above, the present inventor has conducted intensive studies, and has found that a new islet that is easy to handle when transplanting pancreatic islet cells and that can be immediately removed when a defect occurs in the once transplanted pancreatic islet cells. A subcutaneous implantation site was found. Subcutaneous transplantation has previously been reported to require as much as 5,000 islet cells from more than five donor rats to treat diabetes in one diabetic recipient rat (15). There are reports that the landing efficiency is very low.

  Therefore, the present inventor reasoned that such a low engraftment efficiency may be caused by the lack of nutrient vessels, resulting in islet cells dying from hypoxia after transplantation. Then, since the feeding vessels of the inferior abdominal wall artery and vein pass through both sides of the inguinal subcutaneous white adipose tissue (Non-patent Document 16), the inguinal subcutaneous white adipose tissue was selected as a new subcutaneous transplantation site. Implantation efficiency of pancreatic β cells was found to be improved (Patent Document 2).

  However, surprisingly, the present inventor has found that pancreatic β-cells have been created in a subcutaneous white adipose tissue in the groin where pancreatic β-cells have been transplanted, in the vicinity of the transplant site. The fact that pancreatic β-cells not derived from the donor but from the recipient were found in tissues other than the pancreas is extremely revolutionary.

JP 2008-189574 A WO 2015/159889

Danaei, G., et al. Lancet, 2011, 378 (9785): 31-40 Atkinson, M.A., et al. N. Engl. J. Med. 1994; 331: 1428-1436 Ricordi, C., et.al., 2004, Nat. Rev. Imunol., 4: 259-268. Shapiro, A.M., 2000, N. Engl. J. Med., 343: 230-238 Ryan, E.A., et.al., 2002, Diabetes, 51: 2148-2157 Rabinovitch, A., 1993., Diabetes Rev., 1: 215-240 McCall, M., et al. Cold Spring Herb. Perspect. Med. 2, a007823, 2012 Chentoufi, A.A., et al., Clin. Develop. Immun., Vol. 2011, Article ID 103738, 2 pages, 2011; Pepper, A.R., et al., Clin. Develop. Immun., Vol. 2013, Article ID 352315, 1-13 pages Merani, S., et al., British Journal of Surgery, vol. 95, no.12, pp. 1449-1461, 2008 Komori, J., et al. Nat.Biotechnol. 30, 976-983 (2012) Kakabadze, Z., et al. The American Society of Transplantation and the American Society of Transplant Surgeons 13, 2550-2557 (2013) Unger, R.H. and Orci, L. Lancet. 1975; 1 (7897): 14-16 Bramswig, N.C., et al. The Journal of Clinical Investigation, Vol. 123, No. 3, pp. 1275-1284, 2013 Kawakami, Y., Pancreas, 23, 375-381 (2001) Vitali, A., et al., J. Lipid Res. 53, 619-629 (2012)

  Therefore, the present inventor has transplanted the isolated pancreatic islets into subcutaneous white adipose tissue, thereby creating pancreatic β cells, which are novel insulin-producing cells from the recipient, in the subcutaneous white adipose tissue near the transplanted islets. The inventors have completed the present invention.

  Accordingly, an object of the present invention is to provide a method for creating insulin-producing cells, which comprises creating pancreatic β cells, which are insulin-producing cells, in subcutaneous white adipose tissue.

  In a preferred embodiment of the present invention, a new insulin-producing cell is created in the subcutaneous white adipose tissue by allowing foreign insulin-producing cells to coexist in the subcutaneous white adipose tissue in which the vegetative blood vessels are widely distributed. An object of the present invention is to provide a method for creating an insulin-producing cell.

  In a further preferred aspect of the present invention, insulin-producing cells are transplanted into subcutaneous white adipose tissue in which vegetative blood vessels are widely distributed, so that new insulin is produced at a site adjacent to the transplant site in the subcutaneous white adipose tissue. An object of the present invention is to provide a method for creating an insulin producing cell, which comprises creating a producing cell.

  As used herein, the term “subcutaneous white adipose tissue” refers to subcutaneous white adipose tissue of a diabetic recipient, which is localized in the body and can supply essential nutrients to the islet cells. It refers to the subcutaneous white adipose tissue in which the supply blood vessels are widely circulated and distributed in the area. Examples of such subcutaneous white adipose tissue include, but are not limited to, subcutaneous white adipose tissue such as the groin, axillary, back, or abdominal region where the inferior abdominal wall vein circulates. Of these tissues, the groin is preferred because the blood vessels are distributed throughout the area.

  As used herein, the term "vegetative blood vessel" is intended to mean a blood vessel capable of supplying subcutaneous white adipose tissue with nutrients essential for transplanted islets. Such blood vessels can include, for example, arteries, veins, and / or capillaries.

  As used herein, the term "exogenous insulin-producing cells" refers to insulin-producing cells that have a different origin from the cells that make up the coexisting subcutaneous white adipose tissue. This means, for example, that the exogenous insulin-producing cells are derived from the donor and the subcutaneous white adipose tissue is derived from the recipient. In addition, the exogenous insulin-producing cells are not limited whether they are derived from the same species or different species.

  As used herein, the term "coexistence" refers to a state in which exogenous insulin-producing cells coexist in subcutaneous white adipose tissue. As a method in which exogenous insulin producing cells coexist in subcutaneous white adipose tissue, for example, there is a method of transplanting insulin producing cells into subcutaneous white adipose tissue of a diabetic recipient. In order for exogenous insulin-producing cells to coexist in subcutaneous white adipose tissue, it is necessary that the subcutaneous white adipose tissue and exogenous insulin-producing cells are in a co-nutritive state, that is, have a nutrient supply for coexistence.

  In order to achieve the above object, the present invention provides a novel insulin-producing cell in a subcutaneous white adipose tissue by allowing exogenous insulin-producing cells to coexist in a subcutaneous white adipose tissue in which vegetative blood vessels are widely distributed. And a method for creating an insulin-producing cell.

  As a more preferred embodiment for achieving the above object, the present invention provides a method for coexisting exogenous insulin-producing cells with subcutaneous white adipose tissue in which vegetative blood vessels are widely distributed, preferably subcutaneous white adipose tissue in the groin by transplantation. And a method for creating insulin-producing cells, which comprises producing new insulin-producing cells at a site adjacent to a coexisting site in the subcutaneous white adipose tissue.

  The present invention creates new insulin-producing cells in the subcutaneous white adipose tissue of diabetic recipients, for example, in the subcutaneous white adipose tissue where vegetative blood vessels such as groin, axillary, and back are widely distributed. Since it can be expected to reduce the number of insulin-producing cells transplanted from a single donor, it has a great advantage in that it can help alleviate the major problem of pancreatic islet cell transplantation due to a shortage of donors. Furthermore, if a recipient-derived insulin cell-inducing factor is found, insulin-producing cells can be created only by administering the factor into subcutaneous adipose tissue, leading to the development of a revolutionary treatment for diabetes that does not require islet transplantation.

FIG. 1 is a view showing a part of a subcutaneous white adipose tissue of a mouse inguinal region as a new islet transplantation site. FIG. 1 (A) shows the subcutaneous white adipose tissue in the left groin when lifted with forceps. 1A and 1B show the vegetative blood vessels of the subcutaneous white adipose tissue in the groin. Long and short arrows indicate the femoral artery and vein and the lower abdominal wall artery and vein, respectively. FIG. 2 is a diagram showing a technique for transplanting pancreatic islets into subcutaneous white adipose tissue of the mouse groin. FIG. 2a is a top view of the left inguinal skin incision. FIG. 2b shows a small pocket created in the inguinal subcutaneous white adipose tissue. Figures 2c and 2d show the islets attached to the tip of the tube placed inside the pocket. In the drawings, arrows indicate islets in the pockets, and arrowheads indicate islets at the tip of the tube. FIG. 2e shows the pocket opening held by tweezers. FIG. 2f shows the pocket opening closed with a stapler. The scale is 1 mm. FIG. 3 is an explanatory diagram showing changes in insulin content over time in adipose tissue including transplanted pancreatic islets. FIG. 4A is an explanatory diagram showing the results of Western blot of subcutaneous white adipose tissue transplanted with isolated mouse islet cells. FIG. 4B is a diagram showing the ratio of the expression level to an endogenous marker (β-actin). FIG. 5 is a diagram showing an immunostained tissue image of GFP / insulin / cells in adipose tissue of a recipient 120 days after transplantation of mouse isolated islet cells. FIG. 6 is a diagram showing a comprehensive analysis of messages in adipose tissue around transplanted islet cells by RNAseq.

  The present invention relates to a method for transplanting exogenous insulin-producing cells into subcutaneous white adipose tissue of a diabetic recipient and coexisting with the same, thereby producing new insulin-producing cells at a site close to the transplant site in the subcutaneous white adipose tissue into which the insulin-producing cells have been transplanted. And a method for creating an insulin-producing cell.

  In the present invention, the coexisting site of exogenous insulin-producing cells is a subcutaneous white adipose tissue site that can be used as a scaffold for engrafting insulin-secreting cells and secreting insulin among various body parts where subcutaneous white adipose tissue is localized. And any of the subcutaneous white adipose tissue sites, as long as the nutritional supply vessels necessary to supply the foreign insulin-producing cells to the subcutaneous white adipose tissue are circulated and distributed in the tissue. Good. Preferred examples of such a coexisting site include the groin, armpit, back, and abdomen. Of these tissues, the inguinal region is preferred because the circulating lower abdominal wall arteries and veins can supply nutrients to the transplanted islet cells.

  Examples of the foreign insulin-producing cells used in the present invention include human pancreatic islet cells, heterologous pancreatic islet cells such as pigs, and insulin-producing cells created from ES cells and iPS cells.

  Further, in the present invention, as a transplantation method, which is one method of coexisting exogenous insulin-producing cells in subcutaneous white adipose tissue, for example, a cell-encapsulated container in which insulin-producing cells of a donor are encapsulated in a capsule or the like is used for a diabetic recipient. Examples of such methods include embedding in subcutaneous white adipose tissue of the body, or directly transplanting insulin-producing cells for transplantation from a donor into subcutaneous white adipose tissue of a diabetic recipient by injection or the like. In the case of implanting a cell-encapsulated container such as a capsule in which insulin-producing cells are encapsulated, there is an advantage that once the transplanted insulin-producing cells cause transplant rejection, the container can be removed.

  The present invention will be described in detail with reference to examples, but the following examples do not limit the present invention in any way, and are described for illustrative purposes only. Therefore, the present invention includes all modifications and improvements deriving from the description of the above specification and the following examples within the scope of the present invention.

[Materials and methods]
(mouse)
C57BL / 6 was used as a donor mouse. Mice were housed in specific sterile rooms and used for experiments at the age of 8-16 weeks. All experiments were performed according to a protocol approved by the Fukuoka University Animal Care and Use Committee.

  Pancreatic islets were obtained according to the method described in the literature (Sutton, R., et al., Transplantation, 42, 689-691 (1986); Okeda, T., et al., Endocrinol. Jpn. 26, 495-499 (1979)). Isolated. The isolated islets were cultured in a CO2 incubator (5% CO2 + 95% air) using a 10% FBS-supplemented medium (D-MEM, Nissui) at 24 ° C. overnight and used as a donor. The islets were transplanted into the left inguinal subcutaneous white adipose tissue of STZ (Sigma) (180 mg / kg) -induced diabetic syngenic recipient mice on the third day of STZ injection.

  FIG. 1 shows the anatomical findings of the subcutaneous white adipose tissue in the left groin of the mouse. FIGS. 1 (A) and 1 (B) show a state in which the left inguinal subcutaneous white adipose tissue in which a feeding blood vessel is observed is lifted with tweezers (left side), respectively. In the figure, long and short arrows indicate a femoral artery and a lower abdominal wall artery and vein, respectively.

  FIG. 2 shows a state in which pancreatic islets have been transplanted into subcutaneous white adipose tissue of the mouse groin. FIG. 2 (a) is a view of the skin incision of the subcutaneous white adipose tissue in the left groin of the mouse as viewed from above. FIG. 2 (b) shows a small pocket in the groin subcutaneous white adipose tissue, with the opening kept open with sutures. FIGS. 2C and 2D show the islet cells existing at the tip of the tube placed inside the pocket. 2 (c) and 2 (d), the arrows indicate the islet cells arranged in the pockets, and the arrowheads indicate the islet cells at the tip of the tube. FIG. 2E shows the opening of the pocket held by the forceps, and FIG. 2F shows the opening of the pocket closed by the stapler needle. In the figure, the scale is 1 mm.

  The insulin content in adipose tissue including the transplanted islet was measured with time after the islet transplantation. The result is shown in FIG. As shown in FIG. 3, it was found that the insulin content in adipose tissue rapidly decreased 30 days after transplantation, but increased after 60 days and 120 days after transplantation. did.

  In the same manner as in Example 1, a mouse-insulin promotor (MIP) -GFP mouse (C57BL / 6 background) was used as a recipient. The pancreatic β cells of this mouse are labeled with GFP, and if GFP is detected in tissues other than pancreatic islets in the pancreas, it is evidence that it is derived from the recipient. 400 isolated pancreatic islets of two donors from wild-type C57BL / 6 mice were transplanted into the inguinal subcutaneous white adipose tissue of STZ diabetic MIP-GFP recipients. Western blotting using an anti-GFP antibody was performed on adipose tissue including the transplanted islets, and GFP protein was detected 120 days after transplantation (FIG. 4A). Β-actin was used as an endogenous control, and the ratio to the GFP expression level is shown in FIG. 4B.

  Adipose tissue of the recipient STZ diabetic MIP-GFP mouse 120 days after transplantation transplanted in the same manner as in Example 2 was subjected to immunostaining according to a conventional method. FIG. 5 shows the immunostained tissue image.

  The left panel shows immunostained histological images of GFP protein (indicated by an arrow at the upper left) and insulin (lower) of transplanted islets. The upper middle panel shows an enlarged immunostained tissue image of the GFP protein in the left panel, the upper right panel shows an immunostained tissue image of insulin, and the lower lower panel shows a nuclear immunostained tissue image of the cells with DAPI. The lower right figure shows an image obtained by combining these three immunostained tissue images. From this figure, the immunostained tissue images of GFP and insulin overlap and are present adjacent to the transplanted cells. I understand that there is. This result indicates that insulin-producing cells derived from the recipient are newly created in the vicinity of the transplant site of the transplanted islet.

  Extensive analysis of the message was performed by RNAseq according to a standard method in the portion where the immunostained histological images of GFP and insulin confirmed in Example 3 overlapped, that is, the adipose tissue around the transplanted pancreatic islet on the 60th and 120th days after transplantation. . As a result, as shown in FIG. 6, it was found that β-cell related messages such as insulin I, insulin II, Pdx1, MafA, Nkx6.1, and Ngn3 were amplified.

  The present invention relates to the production of novel insulin-producing cells derived from recipients in subcutaneous adipocytes after allogeneic islet transplantation. It is to provide a so-called in vivo reprogramming method for inducing a novel pancreatic β-cell that newly produces insulin-producing cells in white adipose tissue. That is, the present invention provides a novel strategy for treating diabetes. It can also be said that the present invention provides a pancreatic β-cell expression agent for expressing recipient-derived pancreatic β cells in subcutaneous white adipose tissue.

Claims (3)

  Insulin-producing cells, wherein exogenous insulin-producing cells coexist in subcutaneous white adipose tissue in which vegetative blood vessels are widely distributed, thereby producing new insulin-producing cells in the subcutaneous white adipose tissue. Raw method.   The method according to claim 1, wherein the exogenous insulin-producing cells coexist with the subcutaneous white adipose tissue in which the vegetative blood vessels are widely distributed, whereby a coexistence site in the subcutaneous white adipose tissue is formed. A new insulin-producing cell at a site adjacent to the cell.   The method for creating insulin-producing cells according to claim 1 or 2, wherein the coexistence is performed by transplanting exogenous insulin-producing cells into subcutaneous white adipose tissue of a recipient. .