Method for treating or preventing pancreatic dysfunction

Abstract

To provide a method for improving pancreatic function.SOLUTION: The present invention provides a method for improving pancreatic function in a subject in need thereof, the method comprising administering to the subject STRO-1+ cells and / or progeny cells thereof and / or soluble factors derived therefrom. The method of the invention is useful for treating and / or preventing and / or delaying the onset or progression of a disorder resulting from or associated with pancreatic dysfunction, e.g., resulting from abnormal endocrine or exocrine function of the pancreas.SELECTED DRAWING: None

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to a method for improving pancreatic function in a subject in need thereof. , disorders resulting from or associated with pancreatic dysfunction, e.g., pancreatic endothelial Treatment and / or prevention of disorders resulting from abnormalities in secretory or exocrine functions and / or delaying the onset or progression of the disease. [Background technology]

[0002] Background of the Invention The pancreas is a multifunctional glandular organ in the digestive and endocrine systems of vertebrates. Endocrine glands (which produce several hormones, including insulin, glucagon, and somatostatin) It is both an exocrine gland (secreting pancreatic juices containing digestive enzymes that pass through the small intestine) and an exocrine gland (secreting pancreatic juices that contain digestive enzymes that pass through the small intestine) Enzymes in the pancreatic juice help in the further breakdown of carbohydrates, proteins, and fats in the chyme. Standing there.

[0003] The pancreas, which has endocrine functions, is made up of numerous cell clusters called the islets of Langerhans. The islets contain four major cell types classified by secretion: α cells secrete glucagon, β cells secrete insulin, and δ cells secrete somatostatin. PP cells secrete pancreatic polypeptide. The islets are arranged in clusters and cords. The islets are dense collections of endocrine cells that are densely packed together and also contain a network of capillaries. The capillaries are lined with a layer of endocrine cells that directly contact the blood vessels, and most of the endocrine cells Secretory cells are in direct contact with blood vessels either by cytoplasmic processes or by direct apposition.

[0004] In contrast to the endocrine pancreas, which secretes hormones into the blood, the exocrine pancreas secretes digestive enzymes ( For example, trypsinogen, chymotrypsinogen, elastase, carboxypeptides It produces enzymes (lipase, pancreatic lipase and amylase) and alkaline fluid, and is a hormone of the small intestine. In response to secretin and cholecystokinin, which are secreted by the exocrine system, they are distributed to the small intestine. Digestive enzymes are produced and secreted by the acinar cells of the exocrine pancreas. Specialized cells lining the pancreatic ducts, called centroacinar cells, secrete bicarbonate and salts It secretes a rich solution into the small intestine.

[0005] Pancreatic dysfunction is caused by overproduction or over-production of hormones and / or enzymes produced by the pancreas. associated with or caused by pancreatic dysfunction. Conditions that may be present include diabetes mellitus, acute or chronic pancreatitis, and pancreatic enzyme deficiency. or pancreatic tumors.

[0006] Diabetes mellitus (DM) is the most common chronic endocrine disorder across all age groups and populations. DM is a type of diabetes mellitus (DM) caused by pancreatic dysfunction. It affects over 100 million people worldwide. In the United States alone, over 12 million people are diagnosed with DM each year. 600,000 new cases are diagnosed.

[0007] DM is a disorder of carbohydrate (e.g., glucose) homeostasis or metabolism that results in elevated blood glucose. It is a diagnostic term for a group of disorders characterized by abnormalities in the The various components of DM include metabolic, vascular, and neuropathic components that affect the endocrine system of the pancreas. These may be due to metabolic and / or exocrine dysfunction, typically characterized by hyperglycemia. The factors are the absence or significant reduction of hormone secretion, especially insulin (i.e., endocrine function) Changes in carbohydrate, fat and protein metabolism caused by At the exocrine level, the pancreas produces various enzymes involved in the digestion of food. For example, the pancreas produces amylase, which digests carbohydrates in DM. The body is unable to secrete sufficient levels of this enzyme, leading to exocrine pancreatic insufficiency, malnutrition, and This may result in weight loss and impaired endocrine and exocrine functions of the pancreas. The metabolic component of DM contributes to the metabolic component of DM. The vascular component of DM involves cardiovascular, retinal, and renal involvement. Abnormalities of the peripheral and autonomic nervous systems are also components of DM. do.

[0008] DM is generally characterized by a decrease in the amount or circulation of insulin and / or It occurs due to a decrease in cellular response to insulin. Insulin is responsible for the breakdown of carbohydrates, fats, and Insulin is essential for muscle and fat cells in the metabolism of fat. By allowing glucose to enter the body and as a carbohydrate store, It lowers blood sugar levels by stimulating the conversion of glucose into glycogen (glycogenogenesis). Insulin also stimulates the release of stored glucose from liver glycogen (glycogenolysis). ), slowing the breakdown of fat into triglycerides, free fatty acids, and ketones. In addition, insulin stimulates the breakdown of proteins for glucose production (gluconeogenesis). Insulin is produced by beta cells in the islets of Langerhans in the pancreas. , and is secreted.

[0009] Diabetes mellitus includes type 1 (also called insulin-dependent diabetes mellitus or IDDM) and type 2 type 2 diabetes (also called non-insulin dependent diabetes mellitus or NIDDM), gestational diabetes, and diabetes There are several types of diabetes, including pre-morbidity (or glucose metabolism disorders). The two most common forms of diabetes are type 1 and type 2. Type 1 diabetes (or insulin) Diabetes Mellitus (IDDM) is a pancreatic beta cell deficiency that results in a complete lack of insulin. Type II diabetes (non-insulin dependent diabetes mellitus; NID) is caused by the presence, destruction, or loss of insulin. DM) is a heterogeneous disorder characterized by insulin resistance.

[0010] Type I diabetes The overall incidence of type 1 diabetes in the US alone is approximately 15 cases / 100,000 individuals. In the United States, approximately 5 to 15 percent of all cases of diabetes are type 1 diabetes, and Approximately 10,000 new cases are diagnosed each year. Internationally, the incidence of type 1 diabetes is From approximately 0.61 cases / 100,000 individuals in China to approximately 34.5 cases / 100,000 individuals in Sardinia cases / 100,000 and over 40 cases / 100,000 in Finland. Many countries have reported that the incidence of type 1 diabetes has doubled in the last 20 years. Further reports have been made.

[0011] The acute clinical onset of type 1 diabetes is characterized by symptoms such as hyperglycemia, polyuria, polydipsia, weight loss, or visual impairment. Characterized by symptoms, alone or in combination, followed by ketoacidosis days or weeks later Generally, the acute onset of disease is preceded by a long asymptomatic pre-symptomatic period, during which During this time, the insulin-secreting beta cells are thought to be gradually destroyed by the subject's immune system. do.

[0012] In a healthy individual, the pancreas normally contains 1 to 1.5 million islets, and islet cells Approximately 80 percent of the cells are insulin-producing beta cells. Clinical diabetes symptoms are caused by the It occurs when fewer than 10 percent of the beta cells remain.

[0013] The mismatch between insulin supply and demand occurs due to the loss of pancreatic beta cells, leading to the development of glutathione-binding proteins (Gibber). This leads to abnormalities in glucose, lipid, and protein metabolism. Insulin deficiency leads to hyperglycemia. and hyperglycemic dehydration, elevated free fatty acid levels, elevated serum ketone levels, triglycerides Increased cholesterol levels, increased very low density lipoprotein (VLDL) levels, and branched chain amino acids This can lead to increased fatty acid levels, decreased protein synthesis, and ketoacidosis. Subjects with type 2 diabetes may have any one or more of a variety of vascular and neurological complications. For example, people with type 1 diabetes are more likely to have a heart attack. patients were twice as likely to develop gangrene and five times more likely to have complete kidney failure than non-patients. 17 times more likely to develop vision loss and 25 times more likely to lose their sight.

[0014] Treatment / Prevention of Type 1 Diabetes Currently, type 1 diabetes is treated with exogenous insulin, exercise, and dietary management. These forms of treatment do not repair pancreatic damage (i.e., replace destroyed beta-islet cells). It does not replenish the growth factors produced by the beta islet cells, or An attempt is made to avoid necessity.

[0015] The majority of subjects with type 1 diabetes require some form of insulin therapy. At the time, such therapies generally involve monitoring blood glucose and / or insulin levels. and, if necessary, inject recombinant or purified insulin. New forms of insulin that can be administered nasally or orally are also being developed. However, this form of therapy requires continuous monitoring by the subject and Requires at least one dose of insulin daily for the rest of one's life. Failure to do so or administering too much insulin can result in, for example, hyperglycemia, hypoglycemia, or ketoacidosis. There may be a risk of developing rheumatoid arthritis.

[0016] Other compounds currently used to treat type 1 diabetes include, for example, sulfonylureas, biphenylsulfonylureas, These include guanides, α-glucosidase inhibitors, or thiazolidinediones. Each of these compounds has significant drawbacks. For example, sulfonylureas can cause hypoglycemia and It causes hyperinsulinemia, biguanides cause lactic acidosis, and α-glucuronides Cosidase inhibitors cause gastrointestinal side effects, and thiazolidinediones have a slow onset of action. , accompanied by weight gain and the need for frequent liver function tests.

[0017] Glucagon-like peptide-1 (GLP-1) has also been identified as a potential therapeutic agent for diabetes. This peptide is important in pancreatic development, β-cell differentiation, and maintenance of β-cell function. Pancreatic and duodenal homeobox factor-1 (PNF-1), a transcription factor that plays a key role in the regulation of endothelial cell proliferation and endothelial cell death Induce the expression of protease inhibitors of dendritic cell membrane protein 1 (PDX-1) (Babu et al., Mol E 20:3133-3145, 2006). PDX-1 is a G Expression of glucose sensing and metabolism, such as LUT2, glucokinase, and insulin In addition to stimulating insulin expression, GLP-1 also induces pancreatic It has been suggested that it may be a therapeutic agent because it can induce the proliferation of pancreatic beta cells (But eau, Diabetes and Metabolism, vol. 34: pp. S73-S77 However, there are no clinically available drugs that increase the intracellular availability of GLP-1. agents, such as orally active dipeptidyl peptidase-4 (DPPIV) inhibitors or The use of injectable GLP-1 analogues has been limited to the treatment of milder forms of type 2 diabetes. The relatively short half-life of these drugs, the need for their frequent administration, and the severe beta-cell loss The relative lack of efficacy in patients with type 1 diabetes or other insulin-dependent Use as an insulin-sparing agent for oral Even commercially available GLP-1 analogs have short half-lives and require high daily doses. .

[0018] Other treatment options include transplantation of pancreatic islets of Langerhans, which allows insulin It has been shown to reduce dependency (Shapiro et al., New Eng. J. Med. 343:230-238, 2000). However, the application of this treatment is donor-dependent. Constrained by the very limited availability of primary human islets to date The donor must have a beating heart to ensure cell survival during transplantation. (Burns et al., J. Endocrinology, 103:437-443 Page, 2004).

[0019] Stem cells, such as embryonic stem (ES) cells, can also be used to generate therapeutically relevant insulin-producing cells. However, insulin-secreting β cells have been proposed as a suitable source for the production of 2~4×10 9 Even the predicted required levels of β cells / transplantation were not produced from stem cells. Such cell-based therapies could potentially address the key points that cause hyperinsulinemia or hypoglycemia. The proliferative capacity of the replaced cells is tightly regulated to ensure that the cells do not expand. The transplanted cells must be kept within the normal range and must avoid being destroyed by the recipient's immune system. Furthermore, there is a need to overcome the difficulty of developing ES cell-based therapies. In this case, residual ES cells must be removed to avoid the risk of teratogenesis. .

[0020] Type II diabetes Type 2 diabetes accounts for approximately 90-95% of diabetes cases, with approximately 193,000 cases in the United States alone. 00 deaths per year. Type 2 diabetes is the seventh leading cause of all deaths. In Western societies, type 2 diabetes currently affects 6% of the adult population, with a predicted global The incidence of type 2 diabetes is growing at 6% per year. Despite the presence of certain genetic traits that may contribute to the disease, the incidence of this disease is currently increasing. The main causes are the sedentary lifestyle, diet and The rise of obesity is a major threat to human health. Type 2 diabetes is currently recognized internationally as one of the most serious health threats. is recognized by

[0021] Type 2 diabetes occurs when muscle, fat, and liver cells fail to respond normally to insulin This failure to respond (called insulin resistance) This is called a decrease in the number of insulin receptors in these cells, or a decrease in the signaling within the cells. This may be due to dysfunction of the neural pathways or both. Over time, these cells compensate for this insulin resistance by increasing insulin production. Type II, which means the body is unable to produce enough insulin to maintain normal glucose levels indicates the progression of diabetes (Kahn et al., Am. J. Med. 108:2S-8S), 20 2000).

[0022] Treatment of Type 2 Diabetes Conventional treatments for type 2 diabetes are very limited and require minimal or no treatment to minimize complications. Current treatments focus on attempts to regulate blood glucose levels to delay insulin resistance. Antibiotics (metformin, thiazolidinediones (“TZDs”)) or insulin secretion from beta cells Sulfonylureas target either steroid release or steroid release (sulfonylureas, exenatide). and other compounds that act by depolarizing beta cells, have been shown to increase circulating glucose levels. Current therapy induces insulin secretion independently of insulin, and therefore has the side effect of hypoglycemia. Other side effects of steroids include weight gain, loss of response to therapy over time, gastrointestinal problems, and edema. include.

[0023] One currently approved drug, Januvia (sitagliptin), is an incretin-based Increases blood levels of incretin hormones, which increase insulin secretion may reduce glucagon secretion and have other less well-characterized effects However, Januvia and other dipeptidyl peptidase IV inhibitors also It may also affect tissue levels of other hormones and peptides, The long-term consequences of the action of have not been fully investigated. Furthermore, this compound may increase insulin resistance. Does not address issues related to sex.

[0024] Similar to type 1 diabetes, GLP-1 induces insulin secretion and beta cell proliferation. and restore glucose tolerance in glucose-tolerant beta cells. Due to its ability, it has been suggested that it may be a therapeutic agent for type II diabetes. As such, GLP-1 and its analogs are difficult to detect due to their very short half-lives. The therapeutic potential for these diseases is very limited.

[0025] Methods for treating or preventing or delaying the onset or progression of disorders related to pancreatic function and It is apparent from the foregoing that there is a need in the art for methods of improving pancreatic function and / or function. . Summary of the Invention [Means for solving the problem]

[0026] Summary of the Invention In the research leading up to the present invention, the present inventors have found that the This study sought to determine the effects of specific subsets of mesenchymal progenitor cells (MPCs) on the development of inflammatory bowel disease. The authors demonstrated that pancreatic dysfunction was induced by administering streptozotocin (STZ) to mice. This compound ultimately inhibited cell death and pancreatic function in a well-established model of pancreatic cancer. STZ induces inflammation and immune cell infiltration in the pancreatic islets, leading to pancreatic endocrine dysfunction. function (e.g., decreased insulin production) and exocrine pancreatic function (e.g., amylase production) This model induces dysfunction in both the metabolic pathways (decreased glucose production) and the metabolic pathways (decreased glucose production). , for example, as a model for type I diabetes or type II diabetes.

[0027] As exemplified herein, the present inventors have demonstrated that STRO-1 + STZ-treated mice Administration of STRO-1 + Compared with STZ-treated mice that did not receive cells, serum iPSCs were significantly higher. The inventors have demonstrated that STR increases insulin levels and decreases blood glucose levels. O-1 + Induction of the number of PDX-1 expressing cells in the pancreas in a subject by the cells. or an increase in the number of pancreatic beta cells and / or islets (e.g., pancreatic The inventors further demonstrated that STRO-1 promotes beta cell regeneration. + Cells , pancreatic alpha cells, by increasing the number of beta cells and / or decreasing the number of alpha cells The inventors further found that STRO restores the ratio of pancreatic beta cells to pancreatic beta cells. -1 + It has further been found that treatment with the cells induces angiogenesis in the pancreas of a subject. These data combined indicate that STRO-1 + cells and / or their progeny and and / or factors secreted therefrom induce or promote pancreatic regeneration, and / or or improve pancreatic function. + Cells or their descendants The cells or factors derived from them may be useful in treating and / or preventing the toxic effects of STZ in the pancreas. These data support the STRO-1 + Cells or The progeny cells or one or more factors derived therefrom may be used to treat or prevent pancreatic dysfunction. or delay in onset or reduction in severity and / or improvement in pancreatic function and / or pancreatic or or inducing regeneration of those cells and / or improving glucose metabolism (e.g., circulating insulin) This shows that it is possible to achieve this by increasing the thoracic acid level.

[0028] The inventors' findings provide a method for treating and / or preventing pancreatic dysfunction, e.g., diabetes, and / or The present invention provides a basis for methods for delaying the onset and / or progression of rheumatoid arthritis.

[0029] Thus, the present invention provides a method for improving pancreatic function in a subject in need thereof. The method comprises administering to the subject STRO-1 + cells and / or their progeny and / or comprises administering soluble factors derived therefrom.

[0030] The present invention also or alternatively provides a method for treating pancreatic dysfunction in a subject (e.g., a subject suffering from pancreatic dysfunction). The present invention provides a method for promoting or inducing pancreatic regeneration, said method comprising administering a STRO- 1 + The cells and / or their progeny and / or soluble factors derived therefrom are tested. For example, this method may involve administering to the body a medicament for the production of new beta cells and Induce or promote the production of blood vessels and / or microvessels.

[0031] The present invention also or alternatively relates to pancreatic beta cells and / or pancreatic islets. The present invention provides a method for inducing or promoting regeneration of STRO-1 islets, the method comprising: +Cells and / or their progeny cells and / or soluble factors derived therefrom to a subject. The method includes the step of:

[0032] The present invention also or alternatively reduces blood glucose levels in a subject, and / or or a method for increasing blood / serum insulin levels, the method comprising administering a 1 + The cells and / or their progeny and / or soluble factors derived therefrom are tested. The method includes administering the compound to the body.

[0033] The present invention also or alternatively provides a method for increasing the number of pancreatic beta cells in a subject. and / or increase the number of pancreatic beta cells relative to pancreatic alpha cells; and and / or methods for decreasing the number of pancreatic alpha cells and / or increasing the number of pancreatic islets. The method provides STRO-1 + cells and / or their progeny and / or and administering soluble factors derived therefrom to a subject.

[0034] The present invention also or alternatively provides a method for treating pancreatic duodenal homeobocosis in the pancreas of a subject. Increase expression of proliferative dextromethorphan-1 (PDX-1) and / or the number of PDX-1-expressing cells and a method for increasing STRO-1. + Cells and / or their progeny The method includes administering the cells and / or soluble factors derived therefrom to a subject.

[0035] The present invention also or alternatively provides a method for detecting arteriogenesis in the pancreas of a subject. The present invention provides a method for inducing or promoting angiogenesis, the method comprising: TRO-1 + cells and / or their progeny and / or soluble factors derived therefrom The method includes administering the compound to a subject.

[0036] The present invention also or alternatively provides a method for increasing the number of pancreatic beta cell precursors in a subject. and methods for inducing or promoting proliferation of pancreatic beta cell precursors. The method is STRO-1 + Cells and / or their progeny and / or derived therefrom The method includes administering to a subject a soluble factor that induces the inflammatory response.

[0037] In one example, the subject suffers from pancreatic dysfunction.

[0038] In one example, pancreatic dysfunction is a dysfunction of the endocrine function of the pancreas and / or the exocrine function of the pancreas. Preferably, the pancreatic dysfunction is related to or caused by pancreatic dysfunction. resulting in decreased pancreatic function (e.g., decreased endocrine pancreatic function or decreased exocrine pancreatic function) or or related thereto.

[0039] In one embodiment of the present invention, the pancreatic dysfunction is associated with or results from a carbohydrate metabolism disorder. These carbohydrate metabolic disorders can lead to endocrine and / or exocrine dysfunction of the pancreas. In one example, impaired carbohydrate metabolism can be due to impaired insulin production by the pancreas. In another example, carbohydrate metabolism disorders are caused by decreased glucagon levels. This is caused by an increase in the number of alpha cells and / or glucagon levels. In another example, a carbohydrate metabolism disorder (increased expression and / or production and / or secretion). The harm occurs due to a decrease in amylase production by the pancreas. In general, carbohydrate metabolism disorders (or pancreatic dysfunction) are not necessarily characterized solely by pancreatic function. For example, carbohydrate metabolism disorders are not caused by insulin. Also characterized by resistance and / or vascular and / or nervous system components In one embodiment of the present invention, the pancreatic dysfunction is diabetes (e.g., type 1 diabetes or Type II diabetes).

[0040] Preferably, the methods of the present invention involve administering an effective amount or a therapeutically or prophylactically effective amount of STRO -1 + cells and / or their progeny and / or soluble factors derived therefrom. In one example, the method includes administering to the subject a medicament for inducing insulin production. for at least about 1 week, or 2 weeks, or 3 weeks, or 4 weeks A sufficient amount of STRO-1 to induce insulin production during + Cells and / or The method includes administering progeny cells of the cells and / or soluble factors derived therefrom.

[0041] In one example, STRO-1 + cells and / or their progeny and / or Soluble factors derived from the soluble factor α are administered directly into the bloodstream of a subject, although administration at other sites is also excluded. Preferably, STRO-1 + cells and / or their progeny and / or Soluble factors derived from STRO-1 are administered systemically. + Cells and / or or their progeny cells and / or soluble factors derived from them are expressed in the aorta, atria of the heart, Intravenous or intraventricular vessels or vessels connecting to the pancreas (e.g., abdominal aorta, superior mesenteric artery, pancreatic tenudin) In a preferred embodiment, the agent is administered intravenously or intra-arterially into the bile duct (the diploic artery or the splenic artery). STRO-1 + cells and / or their progeny and / or soluble proteins derived therefrom The agent may be administered intra-arterially, for example, into the femoral artery or celiac artery, using, for example, a catheter. It is given.

[0042] Alternatively or additionally, STRO-1 + cells and / or their progeny and / or or soluble factors derived therefrom are administered to the pancreas or a portion thereof of the subject.

[0043] In one example, STRO-1 is administered to a subject + STRO-1 cells bri and , and / or express tissue-nonspecific alkaline phosphatase (TNAP). STRO-1 characterized by cell surface markers or their combination + cell Further populations of progeny and / or soluble The factor is administered to cells expressing STRO-1 or STRO-1. bri Cells that are can be derived from cells that express TNAP. Such progeny also express STRO-1. Get or STRO-1 bri and / or may also express TNAP.

[0044] In accordance with an embodiment of the present invention directed to treating or delaying the progression of pancreatic dysfunction, STRO-1 + The cells and / or their progeny and / or soluble factors derived therefrom are involved in the pathogenesis of the disorder. After diagnosis, it is preferably administered, for example, using standard methods known in the art. In those cases where prevention or delay of pancreatic dysfunction is targeted, STRO-1 + cell and / or their progeny cells and / or soluble factors derived therefrom are involved in the clinical presentation of the disorder. Prior to diagnosis, for example, the subject may be diagnosed with impaired glucose tolerance and / or fasting glycemia. ) and / or type 1 diabetes, T-cell and / or B-cell Autoimmune reactions as indicated by proliferation of the population and / or by production of autoantibodies Responses (e.g., cytotoxicity to pancreatic β-islet cells in the development or progression of type 1 diabetes) Increased T cell proliferation and / or autoantibodies against one or more pancreatic beta-islet cell markers It is administered before or simultaneously with the administration of

[0045] Preferably, the methods described herein, according to any of the examples, are directed to the development of pancreatic dysfunction and / or or progression and / or blood glucose and / or blood / serum insulin levels and and / or the number of beta cells and / or the number of alpha cells and / or the number of pancreatic islets and and / or the number of PDX-1-expressing cells and / or the amount of PDX-1 expression and / or blood vessels For example, the method further includes a step of monitoring or detecting the number of The method may include testing glucose tolerance and / or testing fasting blood glucose and / or or measuring the level of a hormone or enzyme produced by the pancreas, and and / or pancreatic samples are obtained to determine beta cell count and / or alpha cell count and / or or the number of pancreatic islets and / or the number of PDX-1 expressing cells and / or the number of PDX-1 expressing cells Such monitoring may further include determining the amount of blood flow and / or the number of blood vessels. The STRO-1 +cells and / or their progeny and / or cells derived therefrom It may prove necessary or desirable to administer a subsequent dose of a soluble factor.

[0046] As will be apparent to one skilled in the art from the preceding paragraph, the present invention may be practiced in accordance with any of the examples described herein. The method is STRO-1 + cells and / or their progeny and / or cells derived therefrom The present invention should not be considered limited to a single administration of the same or a single soluble factor. Multiple doses may be administered either at different sites or via the same or different routes. The present invention expressly encompasses the provision of STRO-1 + cells and / or their progeny and A single administration of the soluble factors derived therefrom is further contemplated.

[0047] In one example, STRO-1 + cells and / or their progeny and / or Soluble factors derived from STRO-1 may be used in compositions such as + Cells and / or progeny cells and / or soluble factors derived therefrom and carriers and / or excipients Suitable carriers and / or excipients will be apparent to those skilled in the art. may be and / or may be described herein.

[0048] Such compositions may contain additional agents useful in the treatment or prevention of carbohydrate metabolism disorders, e.g. , insulin or amylase and / or peptides associated with normal pancreatic function or is a polypeptide (e.g., cholecystokinin octapeptide or somatostatin or Glucagon or trypsinogen or chymotrypsinogen or elastase or carboxypeptidase or pancreatic lipase). Alternatively, or in addition, STRO-1 + The cells or their progeny may be genetically engineered to contain additional factors, e.g., insulin. phospholipase or amylase and / or peptides or polypeptides associated with normal pancreatic function peptides (e.g., cholecystokinin octapeptide or somatostatin or glucagon Trypsinogen or chymotrypsinogen or elastase or carbohydrate The enzymes (peptidase or pancreatic lipase) can be expressed and preferably secreted. .

[0049] The present invention is STRO-1 + cells and / or their progeny and / or derived cells of the resulting soluble factors or compositions containing them, (i) treatment of pancreatic dysfunction; and / or (ii) improvement of pancreatic function; and / or (iii) inducing or promoting the regeneration of pancreatic beta cells and / or pancreatic islets; and / or or (iv) a decrease in blood glucose levels and / or an increase in blood / serum insulin levels; and / or (v) an increase in the number of pancreatic beta cells and / or pancreatic beta versus pancreatic alpha cells Increased number of cells and / or decreased number of pancreatic alpha cells and / or increased number of pancreatic islets addition; and / or (vi) increased expression of pancreatic duodenal homeobox factor-1 (PDX-1) and / or an increase in the number of PDX-1-expressing cells in the pancreas; and / or (vii) inducing or promoting arteriogenesis or angiogenesis in the pancreas; The present invention further provides uses for:

[0050] The present invention relates to the use of STRO-1 in the manufacture of pharmaceuticals. + cells and / or their progeny and and / or soluble factors derived therefrom, (i) treatment of pancreatic dysfunction; and / or (ii) improvement of pancreatic function; and / or (iii) inducing or promoting the regeneration of pancreatic beta cells and / or pancreatic islets; and / or teeth (iv) a decrease in blood glucose levels and / or an increase in blood / serum insulin levels; and / or (v) an increase in the number of pancreatic beta cells and / or pancreatic beta versus pancreatic alpha cells Increased number of cells and / or decreased number of pancreatic alpha cells and / or increased number of pancreatic islets addition; and / or (vi) increased expression of pancreatic duodenal homeobox factor-1 (PDX-1) and / or an increase in the number of PDX-1-expressing cells in the pancreas; and / or (vii) inducing or promoting arteriogenesis or angiogenesis in the pancreas; The present invention further provides uses for:

[0051] The present invention is applicable to a wide range of animals. For example, subjects may be humans, dogs, cats, Preferably, the subject is a mammal, such as a horse, cow, or sheep; preferably, the subject is a human. In another example, the subject is a human. In another example, the subject is a non-human mammal. [Brief explanation of the drawings]

[0052] [Figure 1]Figure 1 is a graphical representation of the effect of STRO-1+ cells on blood glucose levels (BGL) in STZ-induced diabetic NOD / scid mice. Blood glucose levels were determined in diabetic mice injected with STRO-1+ cells (CM) or vehicle (CV) into the left ventricle 10 days after STZ therapy. Blood glucose levels are mean glucose (mM) + / - SE. Student's t-test was performed and significance was determined at p<0.05. [Figure 2] Figure 2 is a graphical representation showing the effect of STRO-1+ cells on blood glucose levels (BGL) in STZ-induced diabetic NOD / scid mice at days 7, 14, and 21 after treatment compared to baseline after 10 days of STZ treatment. Blood glucose levels were determined in diabetic mice injected with vehicle (CV) or STRO-1+ cells (CM) into the left ventricle. Results are expressed as % change in BGL relative to the start of cell therapy on day 10. Student's t-test was performed and significance was determined at p<0.05. [Figure 3] Figure 3 is a graphical representation showing the effect of STRO-1+ cells on insulin levels in STZ-induced diabetic NOD / scid mice 21 days after cell therapy administration. Serum mouse insulin levels were determined in diabetic mice injected with vehicle (CV) or STRO-1+ cells (CM) into the left ventricle. Mouse insulin values ​​are μg / L + / - SE. Student's t-test was performed and significance was determined at p<0.05. [Figure 4A] Figure 4A is a graphical representation showing the effect of intra-arterial STRO-1+ cells on pancreatic microvascular density in STZ-induced diabetic NOD / scid mice 21 days after cell therapy administration. The total number of microvessels stained with anti-smooth muscle actin (SMA) was determined based on the size distribution / cross-sectional area of ​​pancreatic sections. Data are presented as mean + / - sem, with N=8 animals in the vehicle group and N=6 in the STRO-1 therapy group. Student's t-test was performed and significance was determined at p<0.05. [Figure 4B]FIG. 4B is a copy of a photomicrograph (200×) showing microvessels of various diameters stained with mouse anti-smooth muscle actin IgG2a-FITC in pancreatic tissue from mice treated with STRO-1 cells. [Figure 5A] Figure 5A is a graphical representation showing the effect of intra-arterial STRO-1+ cells on pancreatic mRNA profiles in STZ-induced diabetic NOD / scid mice 21 days after cell therapy administration. RNA was extracted from pancreatic tissue from vehicle (CV) and STRO-1 therapy (CM) groups, reverse transcribed, and PCR-amplified for beta cell regeneration-related transcription factors Mafa, Ngn3, and Pdx-1. Total RNA content was normalized to the housekeeping gene beta-actin. Data are presented as mean ± sem, with N=8 animals in the vehicle group and N=6 animals in the STRO-1 therapy group. Student's t-test was performed, with significance at p<0.05. [Figure 5B] Figure 5B is a graphical representation showing the effect of intra-arterial STRO-1+ cells on PDX-1 positive cells in STZ-induced diabetic NOD / scid mice 21 days after cell therapy administration. Pancreatic tissue stained with anti-PDX-1 was analyzed for PDX-1 positive cells / mm2 of islet area. Data are expressed as mean + / - sem, with N=8 animals for vehicle group, N=6 for STRO-1 therapy, and N=3 for untreated control (no STZ). Student's t-test was performed and significance was determined at p<0.05. [Figure 5C] Figure 5C is a copy of a series of photomicrographs (400x) showing antigen-retrieved, formalin-fixed, paraffin-embedded sections stained with mouse anti-PDX-1 (IgG2b) and detected with goat anti-mouse IgG2b-Alexa555 conjugate. [Figure 6A]Figure 6A is a graphical representation showing the effect of intra-arterial STRO-1+ cells on islet characteristics in STZ-induced diabetic NOD / scid mice 21 days after cell therapy. H&E stained pancreatic tissue was analyzed for islet density and normalized to the cross-sectional area examined. Data are presented as mean + / - sem, N=8 animals for vehicle group and N=6 for STRO-1 therapy. Student's t-test was performed and significance was determined at p<0.05. [Figure 6B] Figure 6B is a graphical representation showing the effect of intra-arterial STRO-1+ cells on pancreatic islet characteristics in STZ-induced diabetic NOD / scid mice 21 days after cell therapy administration. H&E stained pancreatic tissue was analyzed for mean islet diameter and normalized to the cross-sectional area examined. Data are presented as mean + / - sem, N=8 animals for vehicle group and N=6 for STRO-1 therapy. [Figure 6C] Figure 6C is a graphical representation showing the effect of intra-arterial STRO-1+ cells on pancreatic islet characteristics in STZ-induced diabetic NOD / scid mice 21 days after cell therapy administration. H&E stained pancreatic tissue was analyzed for mean islet area and normalized to the cross-sectional area examined. Data are presented as mean + / - sem, N=8 animals for vehicle group and N=6 for STRO-1 therapy. [Figure 7A] Figure 7A is a graphical representation showing the effect of intra-arterial STRO-1+ cells on islet characteristics in STZ-induced diabetic NOD / scid mice 21 days after cell therapy administration. Pancreatic tissue stained with anti-insulin was analyzed for insulin-positive cells / mm2 of islet area. Data are expressed as mean + / - sem, with N=8 animals for vehicle group, N=6 for STRO-1 therapy, and N=3 for untreated control (no STZ). Student's t-test was performed and significance was determined at p<0.05. [Figure 7B]Figure 7B is a copy of a series of photomicrographs (200x) showing antigen-retrieved, formalin-fixed, paraffin-embedded sections stained with guinea pig anti-insulin and detected with anti-guinea pig IgG-Rhodamine conjugate. Treatment groups are indicated below each photomicrograph. [Figure 7C] Figure 7C is a graphical representation showing the effect of intra-arterial STRO-1+ cells on islet characteristics in STZ-induced diabetic NOD / scid mice 21 days after cell therapy administration. Pancreatic tissue stained with anti-glucagon was analyzed for glucagon-positive cells / mm2 of islet area. Data are expressed as mean + / - sem, with N=8 animals for vehicle group, N=6 for STRO-1 therapy, and N=3 for untreated control (no STZ). Student's t-test was performed and significance was determined at p<0.05. [Figure 7D] Figure 7D is a copy of a series of photomicrographs (200x) showing antigen-retrieved, formalin-fixed, paraffin-embedded sections stained with mouse anti-glucagon and detected with goat anti-mouse IgG-FITC conjugate. Treatment groups are indicated below each photomicrograph. [Figure 7E] Figure 7E is a graphical representation of the number of beta cells within islets as a percentage of total alpha+ beta cells. Data shown were calculated from the number of insulin-positive cells / mm2 of islet area and the number of glucagon-positive cells / mm2 of islet area. Data are expressed as mean + / - sem, with N=8 animals for vehicle group, N=6 for STRO-1 therapy, and N=3 for untreated control (no STZ). Student's t-test was performed and significance was determined at p<0.05. DETAILED DESCRIPTION OF THE INVENTION

[0053] Detailed Description of the Preferred Embodiments General Techniques and Selected Definitions Throughout this specification, unless otherwise stated or the context requires, a step A composition of matter, a group of steps or a group of compositions may be one or more. A number (i.e., one or more) of those steps, compositions, groups of steps, or groups of compositions should be interpreted as inclusive.

[0054] Each and every individual embodiment or example described herein is intended to be illustrative and not restrictive, unless otherwise specified. This applies mutatis mutandis to other embodiments of the present invention, for example, the detection of pancreatic dysfunction in a subject. Targeted at treatment and / or prevention and / or delay of onset and / or delay of progression It should be understood that individual embodiments or examples described herein may be used interchangeably with those embodiments expressly set forth herein. For improving pancreatic function and / or inducing or promoting pancreatic regeneration, as listed The method is applied mutatis mutandis.

[0055] Individual embodiments described herein relating to the treatment of pancreatic dysfunction are intended to be illustrative and not restrictive. as if expressly recited in the specification, mutatis mutandis for the treatment of carbohydrate metabolism disorders. should be interpreted as being used.

[0056] Individual embodiments described herein relating to the treatment of pancreatic dysfunction are intended to be illustrative and not restrictive. Diabetes, e.g., type I diabetes or type II diabetes, as expressly recited in the specification. These should be interpreted as applied mutatis mutandis to treatment.

[0057] Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It will be understood that the invention includes all such variations and modifications. It should be understood that the present invention does not involve the steps, particularly those mentioned or shown herein. and further including all of the features, compositions and compounds individually or collectively, and or further includes any and all combinations of features or any two or more of the features.

[0058] The scope of the present invention is not limited to the specific embodiments described herein, These are intended for illustrative purposes only. Functionally equivalent products, compositions and methods are not intended to be limiting unless otherwise specified. As described herein, it is clearly within the scope of the present invention.

[0059] This invention does not involve any technical fields related to molecular biology, microbiology, virology, recombinant DNA techniques, or the like, unless otherwise indicated. Excessive synthesis has been achieved using conventional techniques of peptide synthesis in solution, solid phase peptide synthesis and immunology. Such procedures are described, for example, in Sambrook, Fritsch & Maniatis , Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratories, New Yo rk, 2nd Edition (1989), Vols I, II and DI in their entirety;DNA Cloning: A Practical Approach, Vol. I and II (D.N. Glover, ed., 1985), IRL Press, Oxford, full text; Oligonucleotides Synthesis: A Practical Approach (MJ Gait, ed., 1984), IRL Press, Oxford, complete text and especially the article by Gait therein, pp. 1-22; Atkinson et al., pp. 35-81; Sproat et al., pp. 83-1 15; and Wu et al., pp. 135-151; 4. Nucleic Acid Hybridization: A Practical Approach (B. (D. Hames & S.J. Higgins, eds., 1985) IRL Press, Oxford, full text; Immobilized Cells and Enzymes: A Practical Approach (1986) IRL Press, Oxford, full text; Perbal, B ., A Practical Guide to Molecular Cloning(1984);Methods In Enzymology(S.Colowi ck and N. Kaplan, eds., Academic Press, Inc.), entire series; Knowledge database of Ac JFRamalho Ortigao in cess to Virtual Laboratory website (Interactiva, Germany) , “The Chemistry of Peptide Synthesis”; Sakakibara, D., Teichman, J., Lien, E. Land Fenichel, RL (1976).Biochem.Biophys.Res.Commun.73 pp. 336-342; Merrifield, RB( 1963). J. Am. Chem. Soc. 85, pp. 2149-2154; Barany, G. and Merrifield, R.B. (1979). The Peptides (Gross, E. and Meienhofer, J., eds.), vol. 2, pp. 1-284, Academic Press, New York.12. Edited by Wunsch, E. (1974) Synthese von Peptiden in Houben-Weyls Metoden der Or ganischen Chemie (Muler, E., ed.), vol. 15, 4th edition, parts 1 and 2, Thieme, Stuttgart;B odanszky, M. (1984) Principles of Peptide Synthesis, Springer-Verlag, Heidelberg; Bodanszky, M. & Bodanszky, A. (1984) The Practice of Peptide Synthesis, Springer-Ver. lag, Heidelberg; Bodanszky, M. (1985) Int. J. Peptide Protein Res. 25, pp. 449-474; Han Book of Experimental Immunology, Vol. I-IV (eds. D.M. Weir and C.C. Blackwell, 1986 Blackwell Scientific Publications); and Animal Cell Culture: Practical Applied Mathematics Roach, 3rd Edition (ed. John R.W. Masters, 2000), ISBN 0199637970 are.

[0060] Throughout this specification, unless the context otherwise requires, the term "comprises" or Variations such as "comprises" or "comprising" are used in conjunction with the status of the notation. containing a step or element or integer or a group of steps or elements or integers and any other step or element or integer or group of elements or integers It is understood that this does not mean exclusion.

[0061] As used herein, the term "derived from" refers to a specific whole teger) can be obtained from a particular source, but not necessarily directly from that source This should be interpreted as indicating that STRO-1 + cells and / or their progeny With respect to derived soluble factors, the term refers to one or more factors, e.g., STRO-1 + Thin Proteins produced during in vitro culture of cells and / or their progeny , peptides, carbohydrates, etc.

[0062] As used herein, the term "improvement of pancreatic function" refers to the improvement of one or more of the pancreatic functions in a subject. The above pancreatic function is in a subject who has not been treated according to the present invention (preferably is interpreted to mean enhanced compared to the same function (in the subject before treatment) Such terms should be used to describe, for example, individuals who may be suffering from a glucose metabolism disorder. In subjects with or without risk of developing the condition, the level of insulin secretion is increased. This term includes, for example, the improvement of the control of insulin secretion. in subjects with increased levels of glucagon (e.g., as a result of a glucagon-secreting tumor) and / or hypoglycemia, and furthermore, reduced secretion of glucagon in subjects suffering from hypoglycemia. Contains.

[0063] As used herein, the term "pancreatic dysfunction" The term refers to a condition in which the function of one or more pancreases in a subject is comparable to that of a normal and / or healthy individual. The term "pancreatic function" should be interpreted as meaning any pathological condition that differs from the same function in the pancreatic The term "impaired endocrine and / or exocrine pancreatic function" refers to a condition in which the endocrine and / or exocrine function of the pancreas in a subject is impaired by It encompasses pathologies that are enhanced or decreased compared to normal and / or healthy individuals. For example, "pancreatic dysfunction" refers to the production of insulin, glucagon, somatostatin, and pancreatic polypeptide. trypsinogen, chymotrypsinogen, elastase, carboxypeptidase , characterized by or may be related to levels of pancreatic lipase or amylase, or their abnormalities (i.e., increase or decrease). The term "treat a pancreatic disorder" refers to the normalization of pancreatic function (e.g., the treatment of one or more pancreatic disorders). are reduced to more closely resemble their function in normal and / or healthy individuals It will be apparent to one skilled in the art from the foregoing that the present invention encompasses the treatment of a subject in a manner that is effective or enhances the therapeutic effect of the present invention. For example, such treatment may involve the treatment of insulin and / or beta cells and / or In subjects with abnormally reduced islet levels, increased insulin levels and / or This may result in an increase in the number of pancreatic beta cells and / or an increase in the number of pancreatic islets. Therapy may involve, for example, the treatment of glucagon-secreting tumors of the pancreas, e.g., the treatment of glucagon-secreting alpha cells. by a decrease in number and / or a decrease in the expression, production and / or secretion of glucagon This can reduce abnormally elevated glucagon levels. The meaning of the term "prevention or delay of a disorder" will be clear to those skilled in the art based on the foregoing. .

[0064] Pancreatic dysfunction can occur, for example, due to a decrease in the production of digestive enzymes (e.g., lipase or alpha-glucanase) produced by the pancreas. Nutritional impairment as a result of decreased levels of pancreatic enzymes (e.g., α-amylase) and / or decreased production of pancreatic juices May be associated with conditions that result in malabsorption of nutrients (e.g., carbohydrates, lipids, or proteins) These conditions include pancreatitis, pancreatic insufficiency, ficiency), acquired autoimmune deficiency syndrome Preferred examples include cancer, cystic fibrosis, or Zollinger-Ellison syndrome. This condition is caused by a decrease in amylase or lipase produced by the pancreas. of or relating to.

[0065] Pancreatic dysfunction is a condition associated with abnormalities in the use or metabolism of nutrients by a subject, e.g., hypertension. Blood glucose or hypoglycemia, decreased serum amino acid levels, proteinuria, necrolytic migratory erythema may further be associated with conditions that result in necrolytic migratory erythema, or Such conditions include carbohydrate metabolism disorders, such as diabetes. Examples include tumors (e.g., glucagon-secreting tumors that can cause hyperglycemia). Target tumors include glucagonomas.

[0066] As used herein, the term "carbohydrate metabolism disorder" refers to a condition in which a subject has difficulty consuming carbohydrates. Inability to break down or metabolize or ingest or use one or more forms of the compound, or or breaking down or metabolizing or ingesting or using one or more forms of carbohydrates Decreased ability to absorb carbohydrates, generally due to the level of carbohydrate(s) in the bloodstream of the subject. The term "carbohydrate" should be taken to mean any disorder that results in an increase in blood pressure. Disorders of carbohydrate metabolism are caused by an impaired pancreatic production of hormones involved in the breakdown of carbohydrates (e.g., amylase). associated with or caused by a decrease in the production of ATP. carbohydrate metabolism disorders are characterized by a decrease in the pancreatic production of hormones involved in carbohydrate uptake (e.g. associated with or caused by a decrease in insulin production. Carbohydrate metabolism disorders include type 1 diabetes, type 2 diabetes, idiopathic type 1 diabetes (type 1b), early Onset type II diabetes (EOD), young-onset atypical diabetes (YOAD), young-onset adult diabetes Pathology of Modified Ovarian Disease (MODY), nutritional malformation-related diabetes, gestational diabetes, impaired glucose tolerance (IGT), Pathology of abnormal abdominal glucose, metabolic acidosis, ketosis, syndrome X, hyperglycemia, hypoinsulinemia mannosis, insulin resistance, alpha-mannosidosis, beta-mannosidosis, fructose Intolerance, fucosidosis, galactosemia, Leigh's disease, mucolipidosis, mucopolysaccharidoses or any one or more of the above-mentioned complications. Preferably, a carbohydrate metabolism disorder. is diabetes, for example, type I diabetes or type II diabetes.

[0067] Preferably, the subject suffering from diabetes is - Fasting blood glucose of 7 nmol / L or 126 mg / dl or more; - Random blood glucose level of 11.1 nmol / L or 200 mg / dl or higher with symptoms of diabetes (Collect at any time of the day) - ≥ 11.1 nmol / L or 200 mg / dl orally measured at 2-hour intervals Glucose tolerance test (OGTT) values, obtained at 2- or 3-hour time intervals; and other clinically accepted diabetes markers.

[0068] As used herein, the term "effective amount" refers to the amount of a substance that is capable of functioning in a pancreatic tissue prior to administration. Compared to and / or STRO-1 + cells and / or their progeny and / or or soluble factors derived therefrom, compared to subjects not administered STRO-1 + cells and / or their progeny and / or soluble factors derived therefrom are administered and administering to a subject an amount of STRO-1 sufficient to improve pancreatic function. + Cells and / or or their progeny and / or soluble factors derived therefrom. For example, an effective amount of STRO-1 + cells and / or their progeny and and / or soluble factors derived therefrom affect basal or resting glucose levels (blood glucose). Decreased glycemia and / or improved glucose tolerance and / or blood insulin Increased levels and / or increased levels of glucagon in the serum, pancreas or digestive system somatostatin, pancreatic polypeptide, trypsinogen, chymotrypsinogen, Increased levels of estrase, carboxypeptidase, pancreatic lipase, or amylase An effective amount of STRO-1 + cells and / or their progeny and / or Soluble factors derived from these may, for example, penetrate the vasculature surrounding or within the pancreas or its regions. This increases the blood supply to the pancreas or its surrounding areas. The supplier may require such amounts, for example, + cells and / or their progeny and and / or soluble factors derived therefrom and / or specific subjects and / or pancreatic organs It will be noted that the number of patients with autism will vary depending on the type or severity of the disability. Thus, the term refers to a specific quantity, e.g., weight or number of cells or soluble factors. This should not be construed as limiting the invention, but rather the present invention relates to the treatment of pancreatic function in a subject. Any amount of STRO-1 sufficient to improve + cells and / or their progeny and and / or soluble factors derived therefrom. or sufficient STRO-1 to improve pancreatic function + cells and / or their progeny and / or methods for determining the amount of soluble factors derived therefrom will be apparent to those skilled in the art. and / or as described herein. An effective amount does not necessarily mean that the amount is sufficient to treat pancreatic dysfunction. There is no need for treatment or prevention.

[0069] As used herein, the term "therapeutically effective amount" refers to a therapeutically effective amount of a pancreatic or one or more symptoms of a clinical condition that it causes, as a clinical diagnosis of that condition Reduce or inhibit to levels below those observed and tolerated as Sufficient amount of STRO-1 to + cells and / or their progeny and / or For example, a therapeutically effective amount of ST RO-1 + Cells and / or their progeny and / or soluble factors derived therefrom The level of ATP is varied from that observed in diabetic subjects to that observed in presymptomatic subjects (e.g., subjects with impaired glucose tolerance). or impaired resting blood glucose) or levels observed in normal or healthy subjects Additionally, glucose tolerance can be decreased in a subject.

[0070] As used herein, the term "prophylactically effective amount" refers to a dose of a compound or compound that is effective in treating pancreatic disorders. or to prevent or reduce the onset of one or more detectable symptoms of the clinical condition caused by it. A sufficient amount of STRO-1 to inhibit or + cells and / or their progeny and / or or soluble factors derived therefrom. Efficacy of STRO-1 + cells and / or their progeny and / or derived therefrom The soluble factors may be used to treat diabetes in subjects who are clinically diagnosed with diabetes. It can prevent glucose intolerance.

[0071] As used herein, "treat" or "treatment" or The term "treating" refers to the administration of a therapeutically effective amount of soluble factors and / or cells. and at least one clinical condition associated with or caused by pancreatic dysfunction. It should be construed to mean the reduction or inhibition of symptoms of

[0072] As used herein, "prevent" or "preventing" Alternatively, the term "prevention" refers to the administration of a prophylactically effective amount of soluble factors and / or cells. administration of steroids and at least one clinical condition associated with or caused by pancreatic dysfunction The term "progressive" should also be interpreted as meaning the arrest or prevention of the development of one or more symptoms.

[0073] "Delaying the progression of pancreatic dysfunction" means that a treatment reduces the severity of pancreatic dysfunction in a subject. Such a reduction in severity may be achieved, for example, by reducing one or more of the symptoms of pancreatic dysfunction. Complications, such as malabsorption of nutrients, hypoglycemia, hyperglycemia, ketoacidosis, retinopathy, and vitiligo disorders, hypertension, renal failure, coronary artery disease, peripheral vascular disease, neuropathy (e.g., peripheral neuropathy) or autonomic neuropathy) or increased risk of infection. Alternatively, the reduction in the severity of pancreatic dysfunction may be attributed to treatment using the methods of the present invention. Therapeutic treatment (e.g., insulin administration) compared to subjects without the treatment. The condition is characterized by a decrease in the regularity of a subject's therapeutic treatment or requirement for medical treatment. Furthermore, "reducing the progression of pancreatic dysfunction" refers to the use of a compound that reduces the progression of pancreatic dysfunction. one or more measures of pancreatic dysfunction compared with treatment-naive diabetic subjects The delay in the onset of symptoms is

[0074] As used herein, the term "soluble factor" refers to STRO-1 + Cells Any molecule produced by the cell and / or its progeny that is water soluble, e.g., protein Protein, peptide, glycoprotein, glycopeptide, lipoprotein, lipopeptide, carbohydrate Such soluble factors should be interpreted as meaning substances such as those that are present in cells. Such soluble factors may be present in the cells and / or secreted from the cells. and / or purified In one example of the present invention, the soluble factors are the supernatant or the supernatant Therefore, the present invention relates to the administration of one or more soluble factors. Any examples in should be construed as applying mutatis mutandis to the administration of the supernatant.

[0075] As used herein, the term "supernatant" refers to a liquid medium, preferably a liquid Produced after in vitro culture of mesenchymal precursor cells and / or their progeny in culture medium The supernatant usually refers to the non-cellular material that is produced by culturing cells in a medium under appropriate conditions and for a period of time. Culturing the cells and then removing the cellular material by methods such as centrifugation The supernatant may or may not be subjected to further purification steps before administration. In a preferred embodiment, the supernatant is 5 less than 10 4 Less than, more than Preferably 10 3 Contains less than viable cells, and even more preferably contains no viable cells.

[0076] As used herein, the term "normal or healthy individual" is used in the art. and / or any of the methods described herein. It should be taken to mean a subject who does not suffer from pancreatic dysfunction.

[0077] STRO-1 + cells or progeny thereof, supernatants derived therefrom, or one or more Soluble factors of STRO-1 + Cells include bone marrow, blood, dental pulp cells, adipose tissue, skin, spleen, pancreas, brain, and kidney. organs, liver, heart, retina, brain, hair follicles, intestines, lungs, lymph nodes, thymus, bones, ligaments, tendons, skeletal muscles, Cells found in the epidermis and periosteum, and which are germline, e.g., mesoderm and / or endoderm They can differentiate into the germ layers and / or ectoderm.

[0078] In one embodiment, STRO-1+ cells may be derived from tissues including, but not limited to, fat, bone, cartilage, and the like. Multipotent cells that can differentiate into many cell types, including bone, elastic, muscle, and fibrous connective tissue. These cells enter specific lineage-commitment pathways. tment) and differentiation pathways are mediated by mechanical influences and / or endogenous bioactive factors, e.g., growth from local microenvironmental conditions established by factors, cytokines, and / or host tissues STRO-1+ pluripotent cells therefore divide and over time become non- Daughter cells, either stem cells or progenitor cells, which will reversibly differentiate to give rise to phenotypic cells Non-hematopoietic progenitor cells from which cells are derived is.

[0079] In a preferred embodiment, STRO-1 + The cells may be administered to a subject, e.g., a subject to be treated or are related or unrelated subjects (whether of the same or different species) It is concentrated from the sample obtained from the The term or variations thereof refers to a ratio of one particular cell type or a number of particular cell types. The term "cell population" is used herein to describe a cell population in which the proportion of It is used in this way.

[0080] In a preferred embodiment, the cells used in the present invention are TNAP + , VCAM-1 + , THY-1 + , STRO-2 + , CD45 + , CD146 + , 3G5 + or those One or more compounds selected individually or collectively from the group consisting of any combination thereof Expresses a marker.

[0081] "Individually" means that the invention encompasses the listed markers or groups of markers separately. means that individual markers or groups of markers cannot be listed separately herein. However, the appended claims do not limit the scope of the present invention to any such marker or group of markers, whether individually or This means that the two can be defined so as to be separable from each other.

[0082] "Collectively" means that the present invention relates to any number or combination of the listed markers or It is intended to encompass a group of peptides, such as a number or combination of markers or Even if a group of markers is not specifically listed herein, the appended claims do not necessarily encompass such a group. Any such combination or sub-combination, separately and in any Means that it can be defined as divisible from other combinations or groups of markers .

[0083] Preferably, STRO-1 + STRO-1 cells bright (Synonym STRO-1 b ri ) Preferably, STRO-1 bright The cells were further treated with TNAP + , V CAM-1 + , THY-1 + , STRO-2 + and / or CD146 + One of or There are multiple.

[0084] In one example, the mesenchymal progenitor cells are perivascular mesenchymal progenitor cells, as defined in WO2004 / 85630. These are progenitor cells.

[0085] A cell is said to be "positive" for a given marker because the marker is present on the cell surface. Depending on the extent to which the marker is expressed, the level may be low (lo or dim) or high. (bright, bri), where the term is used in cell classification methods. This may be related to the intensity of the fluorescence or other markers used in the experiment. The distinction between dull and bri relates to the markers used for the specific cell population being classified. It will be understood that cells referred to as "negative" for a given marker are necessarily This term refers to the fact that the marker is expressed by the cell, but not completely absent from the cell. This means that the marker is expressed at a very low level relative to the target protein, and when detectably labeled, - produces a very low signal or is not detectable above background levels This means that there is no

[0086] As used herein, the term "bright" refers to a detectably labeled refers to a marker on the cell surface that generates a relatively high signal. Although not all cells express the same target marker protein, "bright" cells express more target marker proteins than other cells in the sample. It has been proposed that these cells express proteins (eg, antigens recognized by STRO-1). For example, when labeled with a FITC-conjugated STRO-1 antibody, fluorescently activated STRO-1 as determined by cell sorting (FACS) analysis bri The cells are non-bright cells (STRO-1 dull / dim ) produces a larger fluorescent signal. The "bright" cells are the least of the most brightly labeled bone marrow mononuclear cells contained in the starting sample. In another example, "bright" cells comprise approximately 0.1% of the total number of cells contained in the starting sample. At least about 0.1%, at least about 0.5% of the most brightly labeled bone marrow mononuclear cells , at least about 1%, at least about 1.5%, or at least about 2%. For example, STRO-1 bright The cells were analyzed using the "background" method, i.e., ST RO-1 ―The cells expressed STRO-1 on the surface, which was 2 logs higher than that of the control cells. In comparison, STRO-1 dim and / or STRO-1 intermedi ate The cells are less than 2 log orders of magnitude higher, usually about 1 log or "background" " have lower STRO-1 surface expression.

[0087] As used herein, the term "TNAP" refers to tissue-nonspecific alkaline phosphatase. The term is intended to encompass all isoforms of ... , liver isoform (LAP), bone isoform (BAP) and kidney isoform In a preferred embodiment, the TNAP is a BAP. In a preferred embodiment, as used herein, TNAP is obtained from the accession number PTA-72. 82 and deposited with the ATCC under the provisions of the Budapest Treaty on December 19, 2005. It refers to a molecule capable of binding to the STRO-3 antibody produced by a hybridoma cell line.

[0088] Furthermore, in a preferred embodiment, STRO-1 + The cells give rise to clonogenic CFU-F It is possible.

[0089] A significant proportion of STRO-1 + Pluripotent cells differentiate into at least two different germ lineages. Non-limiting examples of lineages to which pluripotent cells can be committed include: are hepatocyte progenitors that are multipotent for bone precursor cells, bile duct epithelial cells, and hepatocytes; Neural restricted cells (neural restricted cells) that can generate glial cell precursors that progress to glial cells and astrocytes restricted cells); neural precursors that progress to neurons; precursors of cardiac muscle and cardiomyocytes, Other cell lines include, but are not limited to, glucose-responsive insulin-secreting pancreatic beta cell lines. but not odontoblasts, dentin-producing cells and chondrocytes and the progenitor cells of: retina Skin cells such as pigment epithelial cells, fibroblasts, and keratinocytes, dendritic cells, hair follicle cells, and renal ducts epithelial cells, smooth muscle cells and skeletal muscle cells, testicular progenitors, vascular endothelial cells, tendons, ligaments, cartilage, Adipocytes, fibroblasts, bone marrow matrix, cardiac muscle, smooth muscle, skeletal muscle, pericytes, blood vessels, epithelium, These cells include gliomas, neurons, astrocytes, and oligodendrocytes.

[0090] In another example, STRO-1 + The cells are unable to give rise to hematopoietic cells in culture.

[0091] In one example, cells are obtained from the subject to be treated and transfected using standard techniques. for culturing in vitro and administering to a subject as an autologous or allogeneic composition; Use to obtain supernatant or soluble factors or proliferated cells. In another useful example of the present invention, one or more cells of a human cell line selected from the group consisting of: Cells from a non-human animal (or from another species if the patient is not human) are used.

[0092] The present invention relates to STRO-1 produced by in vitro culture. + Cells and / or the supernatant obtained or derived from its progeny cells (the latter also referred to as proliferating cells) The use of soluble factors is also contemplated. The proliferating cells of the present invention may be grown under various culture conditions (e.g., in the culture medium) or in a manner that allows the growth of the cells. have a wide range of phenotypes depending on factors such as the number and / or type of stimulatory factors, the number of passages, etc. In certain examples, the progeny cells are obtained from about 2, about 3, about 4, about 5, about 6, about 7, about 8, about After about 9 or 10 passages, progeny cells are obtained from the parental population. However, progeny cells may be obtained from the parental population after any number of passages. It may be obtained from the group.

[0093] The progeny cells can be obtained by culturing in any suitable medium. When used herein, the term "medium" includes the components of the environment surrounding a cell. The medium may be a solid, liquid, gas or mixture of phases and materials. and liquid media that do not support cell growth. Media include agar, agarose, gelatin, and Exemplary gaseous media include those in Petri dishes or This includes the gas phase to which cells growing on other solid or semi-solid supports are exposed. The term "cell culture" refers to a material intended for use in cell culture, even if it is not in contact with cells. In other words, a nutrient-rich liquid prepared for bacterial culture is a medium. Powder mixtures that, when mixed with water or other liquids, become suitable for cell culture are referred to as "powdered media." You can call.

[0094] In an example, the progeny cells useful in the methods of the present invention are those labeled with STRO-3 antibodies. TNAP from bone marrow using magnetic beads - STRO-1 + Isolate the cells and then The cells are cultured and grown (an example of suitable culture conditions is Gronthofer et al. See, s et al., Blood 85:929-940, 1995).

[0095] In one example, such expanded cells (progeny) (preferably after at least 5 passages) are TNAP -, CC9 - , HLA class I + , HLA class II - , CD14 - , CD19 - , CD3 - , CD11a - c - , CD31 - , CD86 - , CD34 - and / or CD80 - However, the expression of various markers may vary in the culture medium described herein. Furthermore, cells of these phenotypes can be grown under different culture conditions. A small population of cells (e.g., chromosomes) that do not have this phenotype(s) may be dominant in the population. For example, a very small percentage of proliferating cells may be CC9 - means that there exists In a preferred embodiment, the expanded cells also have the ability to differentiate into different cell types. Also has.

[0096] In one example, the growth medium used to obtain the supernatant or soluble factors or the cells themselves. The cell population should be characterized in that at least 25% of the cells are CC9, more preferably at least 50%. + Yes It includes cells that

[0097] In another example, the enrichment used to obtain the supernatant or soluble factors or the cells themselves A germ cell population is one in which at least 40%, more preferably at least 45%, of the cells are STRO- 1 + The present invention includes a cell in which:

[0098] In a further example, the proliferating cells express LFA-3, THY-1, VCAM-1, ICAM-1, -1, PECAM-1, P-selectin, L-selectin, 3G5, CD49a / CD4 9b / CD29, CD49c / CD29, CD49d / CD29, CD90, CD29, CD18, CD61, integrin beta 6-19, thrombomodulin, CD10, C D13, SCF, PDGF-R, EGF-R, IGF1-R, NGF-R, FGF-R, Leptin-R (STRO-2 = leptin-R), RANKL, STRO-1 bright and CD146 or any combination of these markers, Alternatively, they may express one or more individually selected markers.

[0099] In one example, the progeny cells are cultured using the pluripotent expansion STRO method described in WO2006 / 032092. -1 + Progeny of pluripotent cells (Multipotential Expanded STRO-1 + Multipotential cell)( The STRO-1 gene is defined and / or described as a STRO-1 gene from which progeny can be derived. + Methods for preparing enriched populations of pluripotent cells are described in WO01 / 04268 and WO20 In an in vitro environment, STRO-1 + Pluripotent cells rarely exist in absolutely pure preparations and are generally tissue-specific. They are thought to exist alongside other cells that are transgenic unipotent cells (TSCCs). 4268 has been shown to recover such cells from bone marrow at purity levels of approximately 0.1% to 90%. The population containing the MPCs from which the progeny are derived can be directly recovered from tissue sources or Or alternatively, it may be a population that has already been expanded ex vivo.

[0100] For example, the progeny may be recovered, unexpanded, substantially purified, or otherwise modified. A population of at least about 0.1, 1, 5, 10, 20, 30, 40, 50, 6 STRO-1 containing 0, 70, 80 or 95% + can be obtained from a population of pluripotent cells This level is determined by, for example, TNAP, STRO-1 bright , 3G5 + , VCAM -1, THY-1, CD146 and STRO-2, individually or collectively. by selecting cells that are positive for at least one marker selected for the This can be achieved.

[0101] MEMPS is a marker for STRO-1 bri The test was positive for marker alkali. Freshly harvested STRO-1 cells were negative for alkaline phosphatase (ALP). + In contrast, freshly isolated STRO-1 + Pluripotent cells are , STRO-1 bri In a preferred embodiment of the present invention, the antibody is positive for both ALP and ALP. and at least 15%, 20%, 30%, 40%, 50%, 60%, or 70%, 80%, 90% or 95% STRO-1 bri , ALP - have the phenotype In a further preferred embodiment, the MEMPS expresses Ki67, CD44 and / or CD Positive for one or more of the following markers: 49c / CD29, VLA-3, and α3β1 In an even more preferred embodiment, the MEMP does not exhibit TERT activity and / or are negative for the marker CD18.

[0102] STRO-1+ The starting population of cells may be prepared as described in WO01 / 04268 or WO2004 / 085 Any one or more of the tissue types presented in 630, i.e., bone marrow, dental pulp cells, adipose tissue, They can originate from tissue and skin, or more widely from adipose tissue, teeth, dental pulp, Skin, liver, kidneys, heart, retina, brain, hair follicles, intestines, lungs, spleen, lymph nodes, thymus, pancreas, bones, It can originate from ligaments, bone marrow, tendons and skeletal muscles.

[0103] In the practice of the present invention, the isolation of cells bearing any given cell surface marker can be achieved by a number of Although this can be achieved by different methods, the preferred method is to use a binding agent (e.g., an antibody or binding of the antigen-binding fragments thereof to the associated marker, followed by high levels of binding or It is understood that the results are due to the separation of those that exhibit either low levels of binding or no binding. It will be understood that the most convenient binding agents are antibodies or antibody-based molecules, with high specificity. Therefore, molecules that are monoclonal antibodies or are based on monoclonal antibodies are preferred. Antibodies can be used for both steps, but other agents can also be used, and therefore Ligands for these markers are then used to enrich for cells bearing antibodies or cells lacking antibodies. It can also be used to

[0104] The antibody or ligand may be attached to a solid support to allow for crude separation. or maximize the viability of the recovered fraction. A relatively crude isolate can be obtained. The particular technique used will determine the effectiveness of the isolation, the associated cytotoxicity, ease and speed of performance, and the need for sophisticated equipment and / or technical capabilities. Separation procedures include, but are not limited to, antibody-coated magnetic For bead-based magnetic separation, affinity chromatography, and solid matrices This may involve "panning" using bound antibodies. Techniques that provide accurate separation are limited. Methods for performing FACS are well known to those skilled in the art. It seems that...

[0105] Antibodies to individual markers described herein are commercially available (e.g., STRO- Monoclonal antibodies against 1 are available from R&D Systems, USA. ), available from the ATCC or other depository organization and / or recognized by the art. It can be made using the techniques described above.

[0106] STRO-1 + The method for isolating cells may be, for example, by recognizing high levels of STRO-1 expression. The first step is a solid-phase sorting step, for example using magnetic activated cell sorting (MACS). As described in patent specification WO 01 / 14268, A second sorting step can then follow, if desired, resulting in high levels of progenitor cell expression. This second classification step may involve the use of two or more markers.

[0107] STRO-1 + The method for obtaining cells may involve a first enrichment step using known techniques, followed by: It may also include a step of harvesting the cell source, thus resulting in surgical removal of tissue. The cells comprising the source tissue will then be separated into a so-called single cell suspension. Separation can be achieved by physical and / or enzymatic methods.

[0108] Once proper STRO-1 + Once a cell population is obtained, the cells are cultured by any suitable method. By cultivating or expanding the cells, MEMPs can be obtained.

[0109] In one example, cells are obtained from the subject to be treated and transfected in vitro using standard techniques. for culturing in vitro and administering to a subject as an autologous or allogeneic composition. , supernatant or soluble factors, or proliferated cells. One or more cells of the established human cell line are used to obtain supernatant or soluble factors. In another useful example of the present invention, a non-human animal (or another animal if the patient is not human) is used. Cells from the same species are used to obtain supernatant or soluble factors.

[0110] The present invention is directed to, but not limited to, non-human primate cells, ungulates, dogs, cats, rabbits, and the like. It can be performed using cells from any non-human animal species, including rodent, avian, and fish cells. Primate cells with which the present invention can be practiced include, but are not limited to, chimpanzees, This includes cells from baboons, cynomolgus monkeys, and any other New World or Old World monkeys. Ungulate cells that can be used include, but are not limited to, bovine, porcine, ovine, caprine, and equine cells. Rodent cells with which the present invention can be practiced include, but are not limited to, hamsters, buffalo, and bison. However, it includes cells from mice, rats, guinea pigs, hamsters, and gerbils. Examples of rabbit species that can be irradiated are domestic rabbits, jackrabbits, wild rabbits, and cottontail rabbits. Chickens (Gallus gallus) are also suitable for practicing the present invention. Examples of birds that can be treated are:

[0111] Cells useful in the methods of the invention may be stored prior to use or before obtaining supernatant or soluble factors. Methods and protocols for the preservation and storage of eukaryotic cells, particularly mammalian cells, are available. , known in the art (e.g., Pollard, JW and Walker, JM (1997) Basic C ell Culture Protocols, 2nd edition, Humana Press, Totowa, NJ; Freshney, RI (2000) Cul (See History of Animal Cells, 4th ed., Wiley-Liss, Hoboken, NJ). Any method that maintains the biological activity of cells, e.g., mesenchymal stem / progenitor cells or their progeny. In a preferred embodiment, cells are stored using cryopreservation. It is maintained and stored by

[0112] Genetically modified cells In one example, STRO-1 + The cells and / or their progeny may be, for example, proteins that provide a therapeutic and / or prophylactic effect, e.g., proteins that provide a therapeutic and / or prophylactic effect, e.g., , insulin, glucagon, somatostatin, trypsinogen, chymotrypsinogen , elastase, carboxypeptidase, pancreatic lipase or amylase or vasculoprotein Polypeptides or cells associated with or causing the enhancement of growth of pancreatic cells or cells Genes that cause the cells to express and / or secrete polypeptides associated with differentiation into vascular cells It will be modified.

[0113] Methods for genetically modifying cells will be apparent to those skilled in the art. For example, in cells The nucleic acid to be expressed is operably linked to a promoter to direct expression in the cell. For example, the nucleic acid may be transfected into various cells of a subject using a promoter, e.g., a viral promoter. a motor, such as a CMV promoter (e.g., a CMV-IE promoter) or S Further suitable promoters are known in the art. and should be construed to apply mutatis mutandis to the examples of the present invention.

[0114] Preferably, the nucleic acid is provided in the form of an expression construct. The term "expression construct" refers to a nucleic acid (e.g., a receptor) that is operably linked in a cell. A control gene and / or a counter-selectable reporter gene In the context of the present invention, an expression construct refers to a nucleic acid capable of being expressed in a target gene. plasmids, bacteriophages, phagemids, cosmids, and viral subgenomic fragments. maintain heterologous DNA in fragments or viral genomes or in an expressible format and / or other nucleic acids capable of replication. It should be.

[0115] It will be apparent to those skilled in the art how to construct expression constructs suitable for the practice of this invention. For example, Ausubel et al. (Current Protocols in Molecular Biology. Wiley Interscience) e, ISBN 047 150338, 1987) or Sambrook et al. (Molecular Cloning: : A Laboratory Manual, Cold Spring Harbor Laboratories, New York, 3rd Edition 2001) For example, the individual components of an expression construct can be synthesized from an appropriate template nucleic acid, e.g., PCR, followed by cloning into an appropriate expression construct, such as a plasmid or phagemid. The DNA is amplified using ronning.

[0116] Suitable vectors for such expression constructs are known in the art and / or Suitable expression vectors for the methods of the invention, for example in mammalian cells, are described herein. The vectors include, for example, the pcDNA vector suite (v vector suite vectors, pCI vector suite vectors (Prome ga), pCMV vector suite vectors (Clontech), pM vectors (C lontech), pSI vector (Promega), VP16 vector (Clontech) tech) or pcDNA Vector Suite vectors (Invitrogen) It is Tar.

[0117] Those skilled in the art will be able to find additional vectors and suppliers of such vectors, e.g., Invitrogen You may be aware of companies such as En Corporation, Clontech, or Promega.

[0118] A method for introducing an isolated nucleic acid molecule or a gene construct containing the same into a cell for expression comprises: Those skilled in the art will appreciate that the technique used for a given organism will depend on known successful techniques. Methods for introducing recombinant DNA into cells include microinjection, DEAE-dextrose, and Ran-mediated transfection, liposome-mediated, e.g. Lipofectamine (Gibco, MD, USA) and / or Cellfectin (Gibco, MD, USA) Transfection, PEG-mediated DNA uptake by using , electroporation and, for example, DNA-coated tungsten or was performed by microparticle bombardment using gold particles (Agracetus Inc., WI, USA). Includes the bombardment.

[0119] Alternatively, the expression construct of the present invention is a viral vector. Suitable viral vectors Nucleic acid delivery and its host cell Conventional viral-based systems for integration into the genome include, for example, retroviral vectors. vectors, including lentiviral vectors or adeno-associated viral vectors. Adenoviral vectors are useful for introducing nucleic acids into host cells, which remain episomal. Viral vectors are an efficient and versatile method for gene transfer in target cells and tissues. In addition, the high transduction efficiency is compatible with many different cell types and has been observed in target tissues.

[0120] For example, retroviral vectors have packaging capacity for up to 6–10 kb of foreign sequence. The gene generally contains a cis-acting long terminal repeat (LTR) with minimal cis-acting L The TR is sufficient for replication and packaging of the vector, followed by delivery of the expression construct to the target cell. It is used to integrate into the host and provide long-term expression. The vectors are murine leukemia virus (MuLV) and gibbon ape leukemia virus (GaLV). , simian immunodeficiency virus (SrV), human immunodeficiency virus (HIV) and their combinations (See, e.g., Buchscher et al., J Virol. 56:2731-2739 (1999) 2); Johann et al., J. Virol. 65:1635-1640 (1992); Sommerfelt et al., Virol. 76: 58-59 (1992); 990); Wilson et al., J. Virol. 63:274-2318 (1989); Miller et al., J. Virol. 65:2220-2224 (1991); PCT / US94 / 05700; Miller and Rosman BioTechniques 7:980-990, 1989; Miller, AD Human Gene Therapy 7:5-14, 1990; Scarpa et al., Virology 75:849-852 , 1991; Burns et al., Proc. Natl. Acad. Sci USA 90:8033-8037, 1993).

[0121] Various adeno-associated virus (AAV) vector systems have also been developed for nucleic acid delivery. AAV vectors can be readily constructed using techniques known in the art. For example, U.S. Patent Nos. 5,173,414 and 5,139,941; International Publication Nos. WO92 / 01070 and WO93 / 03769; Lebkowski et al., Molec. Cell. Biol. 5:3988-3996, 1988; Vincent et al. (1990) Vaccines 90(Cold Spring Harbor Laboratory Press);Carter Current Opinion in Biotechnology 5:533-539, 1992; Muzyczka. Current Topics in Microbiol and Immunol. 158:97-1 29, 1992; Kotin, Human Gene Therapy 5:793-801, 1994; Schelling and Smith Gene Therapy 7:165-169, 1994; and Zhou et al., J Exp. Med. 179:1867-1875, 1994 Please refer to 1994.

[0122] Additional viral vectors useful for delivering the expression constructs of the present invention include, for example, viral vectors. Viruses or aphids of the pox family, such as avian poxviruses and avian poxviruses Viral vectors derived from lufavirus or conjugated viral vectors (e.g. For example, those described in Fisher-Hoch et al., Proc. Natl. Acad. Sci. USA 56:317-321, 1989. ) is included.

[0123] Assaying the therapeutic / prophylactic potential of cells and soluble factors Cellular or soluble factors for treating or preventing or delaying the onset or progression of pancreatic dysfunction It will be apparent to one skilled in the art how to determine a child's capacity.

[0124] For example, cellular or soluble factors (e.g., a mixture of factors or a single factor or factors) A fraction (e.g., derived from affinity purification or chromatography) is For example, administering to a subject a time and under conditions sufficient to provide a therapeutic / prophylactic effect. and assessed resting, basal, or fasting glucose levels, and / or Glucose tolerance tests were performed. Such tests were performed using commercially available kits and / or devices. Basal or fasting glucose levels are measured after fasting, e.g. For glucose tolerance testing, subjects are evaluated at about 8 to about 14 hours. The subjects were fasted and then given glucose (e.g., approximately 1.75 grams of glucose / kilogram of body weight) ) and blood sugar levels are assessed approximately 2-3 hours later. According to the Health Organization, fasting plasma glucose is 6.1 mmol / l (100 mg / dl) should be lower than 6.1 and 7.0 mm Fasting levels between 100 and 126 mg / dl (100 and 126 mg / dl) were considered borderline ("fasting hyperglycemia"). Impaired fasting glycemia) with fasting levels repeatedly exceeding 7.0 mmHg Diabetes is diagnosed when the 2-hour glucose level is 1 / 1 (126 mg / dl) or higher. should be lower than 7.8mmol / l (140mg / dl). Levels between 11.1 mmol / L (140 mg / dL) and 11.1 mmol / L (200 mg / dL) A blood glucose level of over 11.1 mmol / L (200 mg / dL) in 2 hours indicates impaired glucose tolerance. The resulting glucose level confirms the diagnosis of diabetes.

[0125] Preferably, the subject suffers from pancreatic dysfunction. For example, the subject is a non-obese diabetic. (NOD) mice (a model of type 1 diabetes) or streptozotocin-treated Certain mice or rats (models of type I and / or type II diabetes; Lukic et al., Developmental Immunol.6:119-128, 1998 and and Arulmozhi et al., Indian J. Pharmacol., 36:217~2 21, 2004), Goto Kakizaki (GK) rats (I model of type I diabetes), New Zealand Obese (NZO) mice (type II Other models of type 1 and / or type 2 diabetes are available, e.g. See, e.g., Rees and Alcolado, Diabet. Med. 22:359-70; It was described in 2005.

[0126] In such models of pancreatic dysfunction, compared with untreated or untreated animals, cells and / or tissues that have reduced basal glucose levels and / or improved glucose tolerance or soluble factors may treat or prevent or delay the onset or progression of pancreatic dysfunction. It is considered highly likely.

[0127] Alternatively, or in addition, insulin levels can be measured in the circulation of a subject, e.g., by measuring the activity of the enzyme The results were evaluated using immunosorbent assay or fluorescent immunosorbent assay. Cellular and / or soluble factors that increase insulin levels may be useful in treating pancreatic dysfunction or It is believed that there is a possibility of preventing or delaying the onset or progression of the disease.

[0128] Kits and assays for determining serum glucagon or somatostatin levels are available. known in the art and / or described in, e.g., Immuno-Biological Laboratories, Inc. or Millipore Corporation It is commercially available from n.

[0129] Alternatively, or in addition, serum levels of amylase may be measured as described by Caraway, Am.J. Colorimetric analysis was performed as described in Clin. Pathol., 32:97-99, 1959. Using or Rinderknecht and Marbach, Clin.Chem Using fluorimetric analysis as described in [Acta], 29:107-110, 1972. Maintain normal serum amylase levels (e.g., 21-101 U / L) The factors or cells that treat or prevent pancreatic dysfunction or delay the onset or progression of pancreatic dysfunction. It is considered highly likely that this is the case.

[0130] Amylase levels can be measured using pancreatic sections or pancreatic tissue obtained by, for example, oroduodenal intubation. These samples can also be used to determine trypsinogen, chymotrypsinogen, and erythrocyte sedimentation. Samples for measuring levels of acetylcholine, elastase, carboxypeptidase, and pancreatic lipase See, e.g., Connon et al., Digestive Diseases and Sciences, 23: 472-475, 1978, reported the pancreatic lipase level in pancreatic juice. An assay for determining the activity of ATP is described.

[0131] The assays described in the preceding paragraphs may be used to assess the success of subjects receiving treatment as described herein according to any of the examples. It is also suitable for continuous monitoring.

[0132] The present invention provides a method for identifying or isolating cells or soluble factors for the treatment of pancreatic dysfunction. It will be apparent to those skilled in the art from the foregoing that said method further provides (i) administering cells or soluble factors to a subject suffering from pancreatic dysfunction, thereby improving the subject's pancreatic function; evaluating (ii) the pancreatic function of the subject in (i) and the subject's lack of prior administration of cells or soluble factors; comparing the pancreatic function of a control subject suffering from pancreatic dysfunction; Including, The improvement in pancreatic function in the subject compared to the control subject is due to the presence of cellular or soluble Indicates that the factor treated pancreatic dysfunction.

[0133] The present invention provides a method for identifying or identifying cellular or soluble factors for preventing or delaying pancreatic dysfunction. Also provided is a method of separating a substrate, the method comprising: (i) administering cells or soluble factors to a subject, and then detecting pancreatic dysfunction in the subject; Inducing (ii) the pancreatic function of the subject in (i) and the subject's lack of prior administration of cells or soluble factors; comparing the pancreatic function of a control subject suffering from pancreatic dysfunction; Including, The improvement in pancreatic function in the subject compared to the control subject may be due to the addition of cellular or soluble factors. prevents or delays the onset of pancreatic dysfunction.

[0134] The cell may be any cell described herein, according to any example.

[0135] cell composition In one embodiment of the present invention, STRO-1 + The cells and / or their progeny are Preferably, such compositions are administered in a pharmaceutically acceptable carrier and and / or excipients.

[0136] The terms "carrier" and "excipient" refer to a vehicle for storing, administering, and / or biosynthesizing an active compound. refers to compositions conventionally used in the art to facilitate biological activity (e.g., , Remington's Pharmaceutical Sciences, 16th edition, Mac Publishing Company (1980 Carriers may also be used to reduce any unwanted side effects of the active compound. Suitable carriers are, for example, stable, eg, incapable of reacting with other components in the carrier. In one example, the carrier is effective in the recipient at therapeutic dosages and concentrations. It does not produce any significant local or systemic adverse effects.

[0137] Carriers suitable for the present invention include conventionally used carriers, such as water, saline, aqueous Dextrose, lactose, Ringer's solution, buffer solutions, hyaluronic acid and glycols Suitable pharmaceutical carriers and excipients include: The agents include starch, cellulose, glucose, lactose, sucrose, gelatin, malt, Rice, wheat flour, chalk, silica gel, magnesium stearate, sodium stearate, Glycerol monostearate, sodium chloride, glycerol, propylene glycol , water, ethanol, etc.

[0138] In another example, the carrier may be, for example, a medium composition in which the cells are grown or in which the cells are suspended. Preferably, such a medium composition does not cause any harmful effects in the subject to which it is administered. Does not induce any effect.

[0139] Preferred carriers and excipients are those that reduce or prevent cell viability and / or pancreatic dysfunction. or does not adversely affect the cells' ability to slow down.

[0140] In one example, the carrier or excipient provides buffering activity and inhibits the absorption of cells and / or soluble factors. maintains the appropriate pH so that it exerts its biological activity, e.g., the carrier or excipient is PBS has minimal interactions with cells and factors. PBS is an attractive carrier or excipient because it is a stable carrier and allows for rapid release of cells and factors. In such cases, the compositions of the present invention are delivered to the bloodstream or to a tissue or tissue surrounding the tissue. or formulated as a liquid for direct application to an area adjacent to the tissue, e.g., by injection. It can be done.

[0141] STRO-1 + The cells and / or their progeny may further be used to identify any host that is compatible with the recipient. and can be incorporated or embedded in a scaffold that degrades into products that are not harmful to the recipient. These scaffolds provide support and support for cells that are transplanted into a recipient subject. Natural and / or synthetic biodegradable scaffolds are examples of such scaffolds. be.

[0142] A variety of different scaffolds are of good use in the practice of the present invention. Preferred scaffolds are those having limited Natural biodegradable scaffolds are not biodegradable but are biodegradable. Suitable scaffolds for cell transplantation include collagen, fibronectin, and laminin scaffolds. Synthetic materials must be able to support a wide range of cell growth and function. The scaffold may be resorbable. Suitable scaffolds are described, for example, in Vacanti et al., J. Ped. Surg. 23:3-9 1988; Cima et al., Biotechnol. Bioeng. 38:145 1991; Vacanti et al., Plast. Reconstr. Sur g. 88:753-9, 1991, including a polyglycolic acid scaffold, or a polyanhydride scaffold. These include synthetic polymers such as polyols, polyorthoesters, and polylactic acid.

[0143] In another example, cells can be administered in a gel scaffold (Upjohn Company ny's Gelfoam, etc.).

[0144] Cell compositions useful for the present invention can be administered alone or in admixture with other cells. Cells that can be administered together with the compositions of the present invention include, but are not limited to, other pluripotent cells. Different types of cells may be used in the treatment of various conditions, including: cells or pluripotent cells or stem cells or bone marrow cells. They can be mixed with the compositions of the present invention immediately or shortly before administration, or they can be mixed with the compositions of the present invention for a short period of time before administration. They can be co-cultured together.

[0145] Preferably, the composition comprises an effective amount or a therapeutically or prophylactically effective amount of cells. For example, this composition contains about 1 x 10 5 STRO-1 + cells / kg to approximately 1 x 10 7 of STRO-1 + cells / kg or approximately 1 x 10 6 STRO-1 + cells / kg to approximately 5 x 1 0 6 STRO-1 + The exact amount of cells to be administered depends on the patient's age, Depends on various factors, including weight and sex and the degree and severity of pancreatic dysfunction .

[0146] In some instances, the cells are unable to exit into the circulation of the subject, but Factors secreted by the cells are contained within a chamber that allows them to enter the circulation. In the method, a soluble factor is secreted into the circulation of a subject by causing cells to secrete the factor into the circulation of the subject. Such chambers may similarly be implanted at a site within a subject, e.g., They can be implanted in or near the pancreas to increase local levels of soluble factors.

[0147] In some examples of the present invention, patients are immunosuppressed prior to initiation of therapy with the cell composition. Therefore, allogeneic or even allogeneic Even if STRO-1 + Transplantation of cells or their progeny is tolerated in some cases. could be.

[0148] However, in other cases, patients may be pharmacologically immunosuppressed before initiating cell therapy. This may be desirable or appropriate. This can be achieved through the use of an agent or by delivering the cells in an encapsulation device. The cells are able to access the nutrients and oxygen required by the cells and the therapeutic factors. The cells are contained in a capsule that is permeable to immune humoral factors and cells, but is impermeable to the immune system. Preferably, the encapsulant is hypoallergenic and readily absorbed into the target tissue. It is easily and stably positioned and provides additional protection to the implanted structure. These and other methods for reducing or eliminating immune responses are known in the art. Alternatively, the cells can be genetically modified to reduce their immunogenicity.

[0149] Composition of soluble factors In one embodiment of the present invention, STRO-1 + Derived from cells and / or derived from progeny cells The supernatant or soluble factors can be, for example, prepared as a composition containing suitable carriers and / or excipients. Preferably, the carrier or excipient is a biologically active agent such as a soluble factor or supernatant. Does not adversely affect the action.

[0150] In one example, the composition is a composition that stabilizes soluble factors or components of the supernatant, e.g., Preferably, the protease inhibitor is not harmful to the subject. There is not a sufficient amount to have an effect.

[0151] Supernatants or soluble factors derived from STRO-1 cells and / or progeny cells The composition comprising the compound may be, for example, a culture medium or a suitable carrier or a buffer solution, such as a phosphate buffered saline solution. The compound may be prepared as a suitable suspension in saline. Suitable carriers are described herein above. In another example, STRO-1 + derived from cells and / or derived from progeny cells The resulting supernatant or suspension containing soluble factors is an oily suspension for injection. The solvent or vehicle may be a fatty oil such as sesame oil or a synthetic fatty acid ester, e.g., oleic acid. Suspensions for injection include ethyl acetate or triglycerides, or liposomes. may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sodium Optionally, the suspension may also contain suitable stabilizers such as sorbitol or dextran. Alternatively, the composition may further contain an agent that increases the solubility of the compound, allowing the preparation of highly concentrated solutions. It can be made into a performance.

[0152] Sterile injectable solutions may be prepared by combining the supernatant or soluble factors in the required amount in an appropriate solvent with one of the above ingredients. or combinations thereof, optionally by incorporating together and then filter sterilizing. It can be prepared.

[0153] Generally, dispersions are prepared by mixing the supernatant or soluble factors with the base dispersion medium and any of the above listed. The required other ingredients are then incorporated into a sterile vehicle containing the required other ingredients. In the case of sterile powders for the preparation of injection solutions, the preferred method of preparation is to use previously filter-sterilized active ingredients. Vacuum drying and freeze drying of the solution of the ingredients plus any additional desired ingredients to obtain powders thereof According to an alternative embodiment of the present invention, the supernatant or soluble factors are dried to enhance their solubility. The compound may be formulated with one or more additional compounds.

[0154] Other exemplary carriers or excipients are described, for example, in Hardman et al. (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro( 2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams and and Wilkins, New York, NY; Avis et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral al Medications, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage F orms:Tablets, Marcel Dekker, NY; Lieberman et al. (eds.) (1990) Pharmaceutical Dosage For ms: Disperse Systems, Marcel Dekker, NY; Weiner and Kotkoskie (2000) Excipient To xicity and Safety, Marcel Dekker, Inc., New York, NY.

[0155] Therapeutic compositions must be sterile and stable under ordinary conditions of manufacture and storage. The object may be a solution, a microemulsion, a liposome or other ordered structure. The carrier may be, for example, water, ethanol, polyol (e.g., glycerol), rolls, propylene glycol and liquid polyethylene glycol) and their A suitable mixture can be a solvent or dispersion medium. By using coatings such as cellulose acetate, and in the case of dispersions, by maintaining the desired particle size; This can be maintained by the use of surfactants and isotonic agents in the compositions, for example, sugars. , polyalcohols such as mannitol, sorbitol or sodium chloride Prolonged absorption of the injectable compositions can be achieved by the use of agents delaying absorption, e.g., monosodium glutamate. This can be achieved by including tearic acid salts and gelatin in the composition. Soluble factors can be administered in a time-release formulation, for example in a composition which includes a slow-release polymer. The active compound may be combined with a carrier that will protect the compound against rapid release, e.g., in an injectable It can be prepared into controlled release formulations, including plant and microencapsulated delivery systems. , biocompatible polymers such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, Collagen, polyorthoesters, polylactic acid and polylactic-polyglycolic acid copolymers (PLG) can be used. Many methods for preparing such formulations are either patented or proprietary. Generally known to those skilled in the art.

[0156] The supernatant or soluble factors may be, for example, in a suitable matrix that provides a slow release of the soluble factors. The drugs may be administered in combination.

[0157] Further components of the composition STRO-1 + Cell-derived supernatant or soluble factor, STRO-1 + Cells or Their progeny may be administered with other beneficial agents or biomolecules (growth factors, nutrients). When administered with other agents, they may be administered in a single pharmaceutical composition or in separate pharmaceutical compositions. simultaneously with or sequentially with other medications (either before or after the administration of other medications) Bioactive factors that can be co-administered include anti-apoptotic agents (e.g., EPO, E PO mimetibody, TPO, IGF-I and IGF-II , HGF, caspase inhibitors); anti-inflammatory agents (e.g., p38MAPK inhibitors, TGF-β inhibitors, statins, IL-6 and IL-1 inhibitors, PEMIROLAST, TRAIL NILAST, REMICADE, SIROLIMUS, and NSAIDs (non-steroidal anti-inflammatory drugs) anti-inflammatory agents; e.g., tepoxalin, tolmetin, suprofen); immunosuppressants / immunomodulators (e.g., calcineurin inhibitors, e.g., cyclosporine, Rolimus; mTOR inhibitors (e.g., Sirolimus, Everolimus); antiproliferative agents (e.g., azathioprine, mycophenolate mofetil); corticosteroids (e.g., prednisolone, hydrocortisone); antibodies, e.g., monoclonal anti-IL- 2R alpha receptor antibodies (e.g., basiliximab, daclizumab), polyclonal anti- T cell antibodies (e.g., antithymocyte globulin (ATG); antilymphocyte globulin (ALG) monoclonal anti-T cell antibody OKT3); antithrombotic agents (e.g., heparin, Derivatives, urokinase, PPack (dextrophenylalanine-proline-arginine thrombin compound, platelet receptor antagonist, antithrombin Antibody to thrombin, antiplatelet receptor antibody, aspirin, dipyridamole, protamine, hirudin , prostaglandin inhibitors and platelet inhibitors); and antioxidants (e.g., probuco Vitamin A, ascorbic acid, tocopherol, coenzyme Q-10, glutathione cysteine, L-cysteine, and N-acetylcysteine) as well as a local anesthetic.

[0158] In one example, the compositions described herein according to any example are useful in the treatment of pancreatic dysfunction or The composition may contain additional agents for prevention, such as biguanides, thiazolidinediones, steroids, sulfonylureas, benzoic acid derivatives, alpha-glucosidase inhibitors, SGLT2 inhibitors Insulin-sensitive drugs and INGAP peptides, dipeptidyl peptidase-IV inhibitors, insulin-sensitive agonists (e.g., PPAR agonists or biguanides), insulin, insulin mimetics Glucagon receptor antagonist, GLP-I, GLP-I mimetic, GLP-I receptor GIP receptor agonists; GIP, GIP mimetics, GIP receptor agonists, PACAP, PAC AP mimetics, PACAP receptor 3 agonists; cholesterol-lowering agents (e.g., HMG- CoA reductase inhibitors, sequestrants, nicotinyl alcohol, nicotinic acid), PPARα / γ dual agonists or anti-obesity compounds.

[0159] In another example, the compositions described herein according to any example may be used to induce the proliferation of progenitor pancreatic cells. Exemplary factors include Wnt, epithelial growth factor receptor 1 (EGFR), and mitogen-activated receptor 2 (MTR). fibroblast growth factor or TGFβ.

[0160] In another example, the composition described herein according to any example may be used to induce the conversion of progenitor cells to vascular cells. Exemplary factors include vascular endothelial growth factor (VEGF), and vascular endothelial growth factor (VEGF). GF), platelet-derived growth factor (PDGF; e.g., PDGF-BB), and FGF.

[0161] In another example, the compositions described herein according to any example may be used to produce tissue-specific unipotent cells. In this regard, the country International Patent Application No. PCT / AU2005 / 001445 discloses TSCC and STRO-1 + It has been demonstrated that administration of cells can result in enhanced proliferation of TSCC. TSCC is a mixture of pancreatic cells, e.g., beta cells, or pancreatic cells, e.g., islets of Langerhans. Administration of such a composition to a subject may, for example, induce the proliferation of beta cells in the islets of Langerhans. In another example, TSCCs are vascular cells. Administration of the composition to a subject results in, for example, increased nutrients being delivered to the pancreas, e.g., pancreatic This may result in increased production of vasculature in the

[0162] medical devices The present invention relates to a method for use in or in a method according to any of the examples described herein. For example, the present invention provides a medical device such as a syringe or a catheter. Iwa STRO-1 + Cells and / or their progeny and / or cells derived therefrom Other suitable delivery devices containing the soluble factors and / or compositions of the present invention are provided. By this, the syringe or catheter may be used in the methods described herein according to any example. It is packaged with instructions for use.

[0163] In another example, the present invention provides STRO-1 +cells and / or their progeny and / or or soluble factors derived therefrom and / or implants containing the compositions of the present invention. Optionally, the implant may be used in the methods described herein according to any of the examples. Suitable implants are described, for example, in the literature, in which: on the above scaffold and STRO-1 + cells and / or their progeny and / or It can be formed using soluble factors derived from

[0164] Mode of administration STRO-1 + Cell-derived supernatant or soluble factor, STRO-1 + Cells or These progeny can be surgically implanted, injected, delivered (using a catheter or syringe), Alternatively, the implant may be inserted directly into the site in need of repair or augmentation, such as the pancreas or blood system of a subject. The drug may be administered directly or indirectly.

[0165] Preferably, STRO-1 + Cell-derived supernatant or soluble factor, STRO-1 + Thin The cells or their progeny are delivered to the bloodstream of the subject. + Cell-derived The resulting supernatant or soluble factor, STRO-1 + The cells or their progeny may be delivered parenterally. Exemplary routes of parenteral administration include, but are not limited to, intraperitoneal, intraventricular, and intracerebral ventricles. Preferably, STRO-1 + supernatant or soluble factors derived from cells, STRO-1 + The cells or their progeny are distributed within the aorta, the ventricles or atria of the heart, and the pancreas. Arterial connection to the connecting vessels, e.g., abdominal aorta, superior mesenteric artery, pancreaticoduodenal artery, or splenic artery In another example, STRO-1 + supernatant or soluble factors derived from cells, STRO-1 + The cells or their progeny are administered into the femoral or celiac artery.

[0166] When cells are delivered to the ventricle or atrium, the cells are administered to the left ventricle or left atrium, and the cells are delivered to the lungs. Rapid delivery is preferred to avoid possible complications.

[0167] Preferably, STRO-1 + Cell-derived supernatant or soluble factor, STRO-1 + Thin The cells or their progeny are then delivered to the delivery site, for example, using a syringe or catheter. It is usually injected through a central line.

[0168] The choice of therapeutic agent dosing regimen depends on the serum or tissue turnover rate of the entity, the severity of symptoms, and the severity of the disease. The dosage regimen depends on several factors, including the level and immunogenicity of the entity. Maximize the amount of therapeutic compound delivered to the patient consistent with an acceptable level of side effects. Thus, the amount of formulation delivered will depend in part on the particular entity and the severity of the condition being treated. do.

[0169] In one example, STRO-1 + Cell-derived supernatant or soluble factor, STRO-1 + The cells or their progeny are delivered as a single bolus dose. -1 + Cell-derived supernatant or soluble factor, STRO-1 + The cells or their progeny , by continuous infusion, or at intervals of, for example, one day, one week, or 1 to 7 times per week The preferred dosing protocol is to administer the maximum dose or significant desired effect. The total weekly dose is determined by the frequency of the dose used to avoid adverse side effects. Determining the appropriate dosage will depend, for example, on the type and activity of the compound. parameters known or suspected to affect treatment or that are expected to affect treatment The dose is determined by the clinician using a meter or factor. Generally, the dose is higher than the optimal dose. Start with a somewhat lower amount and increase in small increments therefrom to the desired level with respect to any negative side effects. or until optimal effect is achieved. Important diagnostic criteria include the criteria for symptoms of diabetes. nothing.

[0170] In accordance with an embodiment of the present invention directed to treating or delaying the progression of pancreatic dysfunction, STRO-1 + The cells and / or their progeny and / or soluble factors derived therefrom are known in the art. using standard methods known in the art and / or methods described herein, e.g., glucose tolerance. It is also preferred that the administration occurs after diagnosis of the disorder.

[0171] In those cases where prevention or delay of pancreatic dysfunction is targeted, STRO-1 + cell and / or their progeny cells and / or soluble factors derived therefrom, e.g., In subjects with impaired glucose tolerance and / or impaired fasting hyperglycemia, administration prior to clinical diagnosis of the disorder and / or in the case of type 1 diabetes, e.g., T cell and / or or by expanding populations of B cells and / or by autoantibodies (e.g., in pancreatic beta islet cells) Cytotoxic T cells against and / or one or more of the following in the development or progression of type 1 diabetes: is an autoantibody produced by the pancreatic islet cell markers (increased autoantibodies against multiple pancreatic islet cell markers). It is preferably administered prior to or simultaneously with the immune response. Methods for determining or predicting the activity will be apparent to those skilled in the art and / or may be used as described herein. For example, autoantibodies against antigens derived from or present on the surface of pancreatic β cells. Detection of autoantibodies indicates an immune response by the subject against the cells. The assay detects islet cell antibodies in the serum of a subject. The method includes contacting the section with serum from a test subject. The immunoglobulins in the serum from the subject then undergo secondary binding to human immunoglobulins. Detected using labeled antibodies. Fluorescent markers are used to detect islet cell antibodies. Suitable methods are described, for example, in Bottazzo et al., Lancet 2:1279-83, 1999 Alternatively, or in addition, the assay may be performed to detect specific antibodies in a subject. It is used for the detection of autoantibodies that bind to antigens. For example, see Brooking et al. in Chim Acta 331:55-59, 2003) showed that GAD65 describes an ELISA-based assay for the detection of autoantibodies. The assay uses low concentrations of GAD antigen on a microtiter plate to detect autologous antigens in the sample. The antibody is captured by adding biotinylated GAD in the liquid phase, which is then captured by the secondary binding site of the autoantibody. The generation of a non-isotopic detectable signal by biotinylated GAD65 is detected. Ann. New York Acad. Sci 1037:10-15, 2004, describes insulin, IA-2 and We have developed an ELISPOT assay useful for detecting the presence of autoantibodies against GAD65 and GAD65. It is stated.

[0172] Methods for monitoring treatment / prophylaxis Methods for monitoring treatment / prophylaxis will be apparent to those skilled in the art based on the disclosure herein. For example, blood glucose and / or insulin levels and / or amylase Protein levels can be measured using methods known in the art and / or described herein. and is evaluated accordingly.

[0173] In another example, a sample (e.g., a biopsy) of the pancreas is obtained after treatment and the beta cells (e.g., insulin-expressing cells) and / or alpha cells (e.g., glucagon-expressing cells) The number of islets and / or PDX-1 expressing cells is determined by, e.g., immunohistochemistry. Use chemical, immunofluorescent, or nucleic acid amplification assays, such as polymerase chain reaction (PCR). A sample of pancreas (e.g., a biopsy) is obtained following treatment. tment and the number of beta cells (eg, cells expressing insulin) and / or alpha cells (eg, cells expressing glucagon) and / or islets and / or PDX-1 expressing c ells, eg, using immunohistochemistry, immunofluorescence or a nucleic acid amp lification assay, e.g., polymerase chain reaction (PCR). As described herein.

[0174] The present invention is further described in the following non-limiting examples. [Example]

[0175] Example 1 STRO-1 + Treatment of diabetic mice with cells 1.1 Materials and Methods Streptozotocin (STZ)-induced diabetes in mice Seven to eight-week-old male immunodeficient NOD / scid mice (NOD.CB17-Prkdc sc id / J;Animal Research Centre, Perth, Australia lia) with 35 mg / kg of the β-cell toxin streptozotocin (STZ; Sigm a-Aldrich, St. Louis, MO) on days 1–4 after a 4-hour morning fast. STZ was administered intraperitoneally (ip) daily in sodium citrate buffer, pH 4.5. The solution was dissolved in 100 ml of PBS and injected within 15 minutes of preparation. Mice were kept under sterile conditions.

[0176] Cell injection and treatment groups Immunomagnetically selected cells derived from banked bone marrow cells Human STRO-1 + The interstitial cells were essentially ino (Methods Mol Biol. 449:45-57, 2008). The cells were cultured and grown as described previously and obtained from Angioblast Systems, USA. Cryopreserved in ProFreeze™-CDM (Lonza, USA) Passage 4, STRO-1 + Thaw 2.5 x 10 stromal cells 6 Cells were injected in 20 10 days after STZ treatment, NOD / scid Mice were injected with a single dose into the left ventricle (arterial route) through the chest wall of anesthetized mice. Control mice received 200 μl of vehicle (7.5% D) via the arterial or venous route. Inject ProFreeze™-CDM (containing MSO and alpha-MEM) Ta.

[0177] Blood glucose and insulin assays After a 4-hour morning fast, a glucometer (Optimum X ceed(TM) Diabetes Monitoring System; Abb (Ott Diagnostics, Victoria, Australia) Blood glucose was assayed in venous blood. Blood insulin was measured at day 32 before the mice were killed. Mouse-specific ELISA was performed on blood obtained by intracardiac puncture of anesthetized mice. SA Kit (Ultrasensitive Mouse Insulin ELISA) The assay was carried out using a 1000 kJ / ml ELISA kit (Mercodia, Uppsala, Sweden).

[0178] Preparation of tissue samples The animals were euthanized by cervical dislocation, and the pancreas was removed, dissected symmetrically, and halves were divided into 10 % neutral formalin, and the others were fixed in Tissue-Tek OCT Compound ( (Sakura Finetek, Torrance, CA) and stored on dry ice. The pancreas was specifically used for most of the analyses in this study. However, other tissues, such as lung, liver, heart, spleen, stomach, intestine / cecum, bladder, testis, and brain, Collected for pathology.

[0179] Histology and immunofluorescence staining of pancreatic tissue For pancreatic histology, formalin-fixed, paraffin-embedded (FFPE) sections were prepared. , stained with hematoxylin and eosin (H&E). Microscope slides FFPE tissue sections (5 μm) mounted on a plate were deparaffinized and then heated in a pressure cooker in citrate buffer. After antigen retrieval, the sections were incubated with 10% normal goat serum. The mouse-specific molecules were detected in pre-tested and antigen-retrieved tissues by blocking for 2 hours. The following antibodies have been demonstrated to detect: guinea pig anti-insulin (1:100; M illipore, USA), mouse anti-glucagon (10 μg / ml; clone K79b B10; AbCAM), mouse anti-PDX-1 (10 μg / ml; clone 267712; A 2-hour primary antibody (R&D Systems) was used for immunofluorescence detection. After incubation, slides were washed with 0.1% normal goat serum / PBS for 5 min. Wash three times and incubate with a species-specific secondary antibody (1:400; goat anti-mouse Alexa Fluor 55 5; Molecular Probe or goat anti-guinea pig rhodamine; Jackso Laboratories or goat anti-mouse IgG1-FITC; AbCAM) The controls were incubated together for an additional 90 minutes at room temperature. Smooth muscle actin (SMA) in pancreatic tissue was stained using mouse anti-SMA-FITCmAb. b (2 mg / ml; clone 1A4; AbCAM) was performed by direct immunofluorescence. did.

[0180] Immunostaining evaluation Slides were viewed under a Zeiss Observer Z1 microscope (Germany). Images were taken using an AxioCam MRm. H&E or insulin Staining was performed using antibodies against glucagon, PDX-1, and SMA, and fluorescent probes were used. The images of the detected pancreatic sections were then scanned using Axio Vision Rel 4.7 software. 5 mm H&E sections from individual experimental animals were used for analysis. The total number of islets was counted, and the size of the islets was analyzed (area and diameter measurements) by image analysis. The cross-sectional area was normalized to the total cross-sectional area of ​​each antibody. Antigen-retrieved 5 μm FFPE sections stained with either PDX-1 or PDX-1 antibodies, respectively. The total number of positively stained cells was counted and the total cross-sectional area or total island surface area was measured, respectively. The distribution of pancreatic microvessels of various diameters was counted and measured by image analysis. All images were taken at a magnification of 2. Analysis was performed at 0–40×.

[0181] RNA analysis by semiquantitative RT-PCR RNA samples from the pancreas of the experimental group were extracted from a total of 100 mm sections from individual frozen tissues. The tissue extracts were extracted in Trizol reagent. pin Mini RNA Isolation Kit(GE Healthcare RNA was purified using a ELISA kit (University of London, UK). Total RNA was quantified spectrophotometrically and 1 μL g is oligo-dT (pdT 12-18 ) and reverse transcribed using MMLV reverse transcriptase Primers for the murine genes for MafA, Ngn3, and Pdx-1 were used. PCR was performed using Tth Plus DNA polymerase (Roc) under the amplification conditions specified in Table 1. cDNA samples were PCR amplified using the Applied Science. The actin gene was used to normalize target gene expression. PCR products were analyzed by UV illumination. Using Kodak ID3.5 software, densitometric analysis of the bands visualized below was performed. and quantified.

[0182] [Table 1]

[0183] statistical analysis Student's T-test was used for P values.

[0184] 1.2 Results Streptozotocin-induced hyperglycemia in NOD / scid mice. In NOD / scid mice, STZ was administered intraperitoneally at a dose of 35 mg / kg / day once daily for 4 days. Hyperglycemia was induced by intraperitoneal injection on day 1 of the study, i.e., before the first STZ injection. The mean fasting blood glucose level (BGL) for all animals (N=80) was 7mM + / - 1.5 On day 10 of the study (i.e., 5 days after the completion of the STZ course), , animals had BGL levels >3 SD above the mean glucose level in untreated mice If an animal had hyperglycemia, it was considered to have developed hyperglycemia. Mice that met the above criteria for hyperglycemia on day 10 were subsequently excluded from all analyses. In these mice, the mean BGL on day 10 was 1 The mean serum creatinine concentration was 5.2mM + / - 0.6, a 217% increase from baseline.

[0185] STRO-1 in diabetic NOD / scid mice + Blood glucose levels after intra-arterial injection of interstitial cells Effect on Value As shown in Figure 1, a single dose of 2.5× 10 6 STRO-1 + The cells also demonstrated a reduction in BGL throughout the 3-week course of cell therapy. It resulted.

[0186] Figure 1 shows that STRO-1 + Single intra-arterial injection of cells This indicates that BGL decreased early in diabetic mice. This was evident as early as day 1 and was greatest at day 24 (mean 35% reduction, mean BGL 12.7mM+ / -1.2 vs 19.6mM+ / -2.1; p=0.012), 3 This persisted throughout the week follow-up period.

[0187] STRO-1 + A single intra-arterial injection of cells resulted in an early and sustained increase in blood glucose levels relative to baseline. resulting in a decrease in value STRO-1 + STZ-treated mice that received a single intra-arterial injection of cells showed no significant changes during the entire 3-week follow-up period. A sustained reduction in mean BGL was achieved over the course of the study compared to baseline levels on Day 10. As shown in Figure 2, this group of animals maintained pre-therapy levels throughout the entire study period. While media-treated controls demonstrated a progressive increase in BGL levels, On the 10th day after STZ treatment, STRO-1 + The groups that received intra-arterial injection of cells were 7 , -11%, -14%, and -4% relative to baseline BGL on days 14 and 21 In contrast, the control group, which received intra-arterial medium alone, demonstrated a mean BGL reduction of 1.0, ... +8%, +20% and +17% vs. baseline BGL on days 7, 14 and 21 demonstrated an increase in the mean BGL of

[0188] Human STRO-1 expression in diabetic NOD / scid mice + A single intra-arterial injection of cells , resulting in a significant increase in circulating levels of insulin in mice after 3 weeks As shown in Figure 3, measured by mouse-specific insulin ELISA at 21 days after treatment Circulating serum insulin levels were measured by STRO-1 + The cells were injected intra-arterially 3 weeks prior. Diabetic mice treated with DM1 had significantly higher circulating endogenous insulin levels compared to vehicle-treated diabetic mice. demonstrated that the urinary tract infections were associated with increased serum urinary tract infections (0.79 mg / L + / - 0.11 vs. 0.57 mg / L). + / -0.02; p=0.009).

[0189] STRO-1 + A single intra-arterial injection of cells significantly increased the pancreatic Increased microvascular density in the liver Pancreatic tissue was incubated with a directly conjugated monoclonal antibody against smooth muscle actin protein. Staining was performed using a mAb (STRO-1) + Cell therapy promotes arteriogenesis in injured pancreas After immunostaining, all sections were scanned and analyzed to determine whether or not the cells induced arteriogenesis. The total number of microvessels was counted and normalized to the total cross-sectional area. Three different vessel diameters were classified based on size: 20-100 μm and >100 μm. Figure 4 shows the STRO-1 + In the cell therapy group, smooth muscle actin-positive microorganisms with a diameter of <20 μm The number of small vessels increased by 176% compared to the vehicle group (299.8 + / - 52 vs. 169 .1+ / -18.5; p=0.01). + Using cells Therapy induces a response of small caliber arterioles within the injured pancreas.

[0190] STRO-1 + A single intra-arterial injection of cells significantly reduced P Increased expression of the DX-1 transcription factor in the pancreas Human STRO-1 + Cell therapy targets endogenous beta cells in diabetic NOD / scid mice To assess whether the regenerative response can be induced in the developing pancreas and beta cells, Expression levels of mRNA for PDX-1, MafA, and Ngn3 transcription factors related to production (Zhou et al., Nature 455:627-632, 2008). As shown in 5A, STRO-1 + In the cell therapy group, there was a significant increase in the number of patients with PDX-1-related transcription factors. Mean pancreatic mRNA levels were increased 2.5-fold compared to the vehicle group (p=0.01). Pancreatic mRNA levels for transcription factors MafA and Ngn3 were also increased, but not significantly. did not reach that level.

[0191] Increased protein levels of PDX transcription factors, such as STRO-1 + Pancreas exposed to cell therapy To confirm that the islet expression was confirmed, healthy non-diabetic NOD / scid mice, Diabetic NOD / scid mice treated with control medium and intra-arterial STRO-1 + Thin Islet sections from diabetic NOD / scid mice treated with vesicles were incubated with anti-PDX-1 As shown in Figure 5B, the mAb was used to immunohistochemically test the streptozotocin. Treatment with rifampin significantly increased the mean number of islet cells that were positive for PDX-1 protein compared with healthy, non-diabetic mice. resulted in a 59% reduction compared to (37.1 + / - 12 mean positive cells / islet vs. 15.1 + / - 4.8 mean positive cells / islet, p=0.03). Compared with syn-treated animals, STRO-1 + Intra-arterial injection of cells results in PDX-1 protein-positive Increased the number of islet cells positive by an average of 71% (25.7 + / - 2.2 mean positive cells / islet, p=0.049), the mean reduction in PDX-1 protein-positive cells compared to non-diabetic animals was The fluorescence micrograph in Figure 5C shows a representative non-diabetic group. Pancreatic islets from target NOD / scid animals or diabetic animals with STRO-1 + Using cells Islets from representative NOD / scid animals treated with either In contrast, a representative diabetic patient receiving control medium demonstrated 1 positive cells. Pancreatic islets from NOD / scid animals showed a significant increase in the number of cells positive for PDX-1 protein. have demonstrated a significant reduction.

[0192] STRO-1 + A single intra-arterial injection of cells results in an increase in islet numbers STRO-1 + The effect of cell treatment on total islet numbers was further evaluated. As shown, 3 weeks ago, STRO-1 + Animals that received a single intra-arterial injection of cells were sacrificed. At this time point, there were more than two-fold more islets (0.78%) than in controls injected with medium alone. + / -0.07 vs. 0.38+ / -0.07 islands / mm 2 , p=0.0012) In addition to an increase in total islet number, there was no significant difference in mean islet diameter or islet area between treatment groups, Figures 6B and 6C. No significant changes were observed.

[0193] STRO-1 + A single intra-arterial injection of cells resulted in endometrial hyperplasia in diabetic NOD / scid mice. Increased number of islet beta cells, decreased number of alpha cells, and normal beta / alpha cells in the islets Reestablishing the ratio Using anti-mouse insulin mAb staining, healthy non-diabetic NOD / scid mice, Diabetic NOD / scid mice treated with control medium and STRO-1 + Using cells The number of beta cells in the islets in the pancreatic slices of diabetic NOD / scid mice treated intra-arterially with α-glucan was increased. As shown in Figure 7A, streptozotocin treatment resulted in the quantification of leukemia in healthy, non-diabetic mice. The number of beta cells in the islets was reduced by 21% (6726 ± 450 / mm3) compared with the control group. 2 Island surface Area ratio 5289+ / -387 / mm 2 , p=0.04). Compared with syn-treated animals, STRO-1 + Intra-arterial injection of cells increases beta cell number by an average of 8% Add (5709+ / -690 / mm 2 ), mean reduction in beta cells compared to non-diabetic animals The decrease was only 15% (p=NS). In the fluorescence micrograph in Figure B, a representative non-diabetic Intraislet beta cells from diseased control animals typically consisted of tightly packed insulin-positive cells in the central region of the islet. However, in a representative STZ-treated mouse, The cells are not abundant and show a segregated pattern within the islets. + Treatment using cells Representative mouse beta cells implanted exhibited an intermediate pattern of fluorescence, which was more abundant and less separated. It proves.

[0194] Anti-glucagon mAb staining was used to identify cells in healthy, non-diabetic NOD / scid mice, control cultures, and Diabetic NOD / scid mice treated with rhesus monkeys and STRO-1 + Cell-based Quantification of intraislet alpha cell numbers in pancreatic sections from intravenously treated diabetic NOD / scid mice As shown in Figure 7C, streptozotocin treatment resulted in a decrease in the number of diabetic mice in healthy non-diabetic mice. In comparison, the number of alpha cells in the islets increased by 470% (1046 + / - 46 / mm 2 Island area 4954+ / -632 / mm 2 , p=0.003). Compared with zotocin-treated animals, STRO-1 + Intra-arterial injection of cells increases average alpha cell numbers 44% reduction (2764 + / - 274 / mm 2 , p=0.008), compared with non-diabetic animals In comparison, the mean increase in alpha cells was only 164% (p=0.002). In fluorescence micrographs, normal islet alpha cells from a representative non-diabetic control animal are organized These can be identified as circumferentially arranged glucagon-stained cells. In mice, alpha cells are more abundant and display a diffuse pattern throughout the islet. STRO-1 + Alpha cells in the islets of a representative mouse treated with cells Demonstrating a pattern where fluorescence is more peripheral and less abundant in the center of the island.

[0195] Figure 7E shows healthy, non-diabetic NOD / scid mice, diabetic mice treated with control medium, and NOD / scid mice and STRO-1 + Diabetic NOD / sc treated intra-arterially with cells The ratio of beta cells to alpha cells within islets of pancreatic sections from id mice is shown. Leptozotocin treatment significantly reduced the number of alpha and beta cells in islets compared to healthy non-diabetic mice. This resulted in a 40% decrease in the percentage of beta cells relative to the total number of cells (8 6+ / -0.9% vs. 52+ / -2.6%, p=0.00002). Compared with streptozotocin-treated animals, STRO-1 + Intra-arterial injection of cells increased the ratio of beta cells to total cells by an average of 29% (52+ / -2.6% vs. 67+ / - -3.9%, p=0.005). Therefore, streptozotocin-induced diabetes STRO-1 in NOD / scid mice + Cell treatment targets alpha cells within the pancreatic islets. This resulted in the re-establishment of a more normal ratio of beta cells to erythrocytes.

[0196] Consideration This study was conducted in streptozotocin-induced diabetic NOD / scid mice. Human STRO-1 + A single injection of cells induces sustained beta cell regeneration and reversal of hyperglycemia. This provides the first evidence that streptozotocin was effective in treating diabetes. The STZ-induced experimental model shows the diabetic features observed after knockdown of the PDX-1 gene. This results in a diabetic phenotype similar to the present-day type, with a reduced number of insulin-producing beta cells and glucagon-producing Increased insulin-producing alpha cells and decreased GLUT2 mRNA and protein expression (Liu et al., Mol Ther 15:86-93, 2007; and Wang et al., Diabetes 47:50-6, 1998). STRO-1 + A single dose of cells , sustained PDX-1 activation, increased endogenous beta cell numbers, and glucagon-expressing alpha cells This resulted in a reduction in blood cholesterol and enhanced insulin production.

[0197] STRO-1 + in STZ-treated NOD / scid diabetic mice after a single injection of cells , sustained induction of PDX-1 expression and homeostasis between pancreatic beta and alpha cells The re-establishment of this effect was confirmed by the long-term administration of glucagon-like peptide-1 (GLP-1) in the same murine model. This is strikingly similar to the characteristics reported after administration of gene therapy to induce L (Iu et al., Mol Ther 15:86-93, 2007). GLP-1 is secreted into the pancreas. A migrating gut-derived peptide that activates PDX-1 and GLUT2 and induces insulin This finding leads to increased GLP-1 activity in beta cells. Two new classes of drugs are: (a) long-acting receptor agonists or GL Dipeptidyl peptidase IV (DPPIV), a natural antagonist of P-1, (2) GLP-1 analogs that are either resistant to degradation by endogenous GLP-1 This has led to the development of an orally active DPPIV antagonist that results in increased GLP-1 activity. .

[0198] However, the clinical use of these drugs is limited to the treatment of mild type II diabetes. Their relatively short half-life, the need for frequent administration, and efficacy in cases of severe beta cell loss The relatively weak insulin resistance makes it difficult to administer insulin to patients with type 1 diabetes or other insulin-dependent individuals. Their use as maintenance drugs is excluded. Indeed, DPPIV antagonists Established diabetes in STZ-treated mice despite increased gonadotropin-dependent GLP-1 levels (Kim et al., Diabetes 50:1562-1570, 2001) ), in the setting of continuous administration of low-dose STZ and partial beta cell loss, improvement of hyperglycemia Only good is possible (Mu et al., Diabetes 55:1695-1704, 200 Similarly, GLP-1 agonists may be given before or simultaneously with STZ. It is only effective if administered continuously and requires continuous administration (Tourrell et al., Diabetes 50:1562-1570). , 2001; Li et al., J Biol Chem 278:471-478, 2003; Gezginci-Oktayoglu and Bolken t, Biochem Cell Biol 87:641-651, 2009). Taken together, these data support the idea that DPP IV inhibitors and GLP-1 analogs are useful for reducing beta cell proliferation when a significant beta cell population is also present. This suggests that the compound is only effective in facilitating tumor cell regeneration.

[0199] In contrast, this study + Even a single injection of cells can result in a large fraction of the beta cell population. These results suggest that the β-cell regeneration pathway can be sustained even in the absence of β-cell proliferation. This was confirmed in a model with complete beta cell loss 5 days after completion of a course of high-dose STZ. This is evidenced by the cells' ability to reverse established hyperglycemia when administered intravenously. Such results can only be achieved by sustained overexpression of GLP-1 using gene therapy. (Liu et al., Mol Ther 15:86-93, 2007). STRO-1 + Long-lasting effectiveness of therapy The potential benefits of this type of cell therapy are comparable to those of DPPIV inhibitors or other treatments in insulin-dependent diabetes. sustained glucose-regulating and insulin-sparing effects not possible with GLP-1 or GLP-1 analogs This shows that it is possible to provide

Claims

1. 1. A method of improving pancreatic function in a subject in need thereof, comprising administering to the subject a STRO -1 ＋ cells and / or their progeny and / or soluble factors derived therefrom. providing a method for detecting a signal;

2. STRO-1 ＋ cells and / or their progeny and / or soluble 10. The method of claim 1, wherein administration of the sex factor promotes regeneration of pancreatic beta cells and / or pancreatic islets. method.

3. STRO-1 ＋ cells and / or their progeny and / or soluble Administration of the sexual factor reduces blood glucose levels and / or increases blood / serum insulin levels in the subject.

3. The method of claim 1 or claim 2, wherein the method increases insulin levels.

4. STRO-1 ＋ cells and / or their progeny and / or soluble administration of the sex factor increases the number of pancreatic beta cells and / or increases the number of pancreatic beta cells in the subject Increase the number of pancreatic beta cells relative to pancreatic alpha cells and / or Any one of claims 1 to 3, which reduces the number of cells and / or increases the number of pancreatic islets.

10. The method according to claim 1.

5. STRO-1 ＋ cells and / or their progeny and / or soluble Administration of sex factors increases the expression of pancreatic duodenal homeobox factor-1 (PDX-1) and / or increase the number of PDX-1-expressing cells in the pancreas of a subject. The method according to any one of claims 1 to 4.

6. STRO-1 ＋ cells and / or their progeny and / or soluble administration of the sex factor induces or promotes arteriogenesis or angiogenesis in the pancreas of the subject. The method according to any one of claims 1 to 5.

7. the subject suffers from a pancreatic dysfunction associated with the endocrine and / or exocrine function of the pancreas; The method of claim 1 or 6.

8. Pancreatic dysfunction may result in the production of insulin, glucagon, somatostatin, pancreatic polypeptide, and triphosphatase. trypsinogen, chymotrypsinogen, elastase, carboxypeptidase, pancreatic lipase 8. The method of claim 7, which is related to or causes abnormalities in the levels of enzymes or amylases. The method described.

9. 9. The method according to claim 8, wherein the abnormality in glucagon levels is caused by a glucagon-secreting tumor. The method described.

10. 8. The method of claim 7, wherein the pancreatic dysfunction is associated with or causes nutrient malabsorption. How to do it.

11. Pancreatic dysfunction can lead to pancreatitis, pancreatic insufficiency, acquired autoimmune deficiency syndrome, cancer, cystic fibrosis, or other conditions. or Zollinger-Ellison syndrome.

12. Pancreatic dysfunction is manifested by hypoglycemia or hyperglycemia, decreased serum amino acid levels, proteinuria, or necrolytic migratory erythema.

13. 8. The method according to claim 7, wherein the pancreatic dysfunction is associated with or causes a carbohydrate metabolism disorder. How to post.

14. Disorders of carbohydrate metabolism can result in decreased insulin production by the pancreas or increased amylase production by the pancreas. The method of claim 7, which is caused by a decrease in production.

15. 15. The method of claim 13 or claim 14, wherein the carbohydrate metabolism disorder is diabetes.

16. STRO-1 ＋ cells and / or their progeny and / or soluble 16. The method of any one of claims 1 to 15, wherein the sex factor is administered directly into the bloodstream of the subject. 。

17. STRO-1 ＋ cells and / or their progeny and / or soluble 17. The method of claim 16, wherein the sex factor is administered intra-arterially.

18. STRO-1 administered to a subject ＋ The cells ｂｒｉ and / or express tissue non-specific alkaline phosphatase (TNAP) and / or progeny cells The cells and / or soluble factors are STRO-1 ｂｒｉ STRO-1 ＋ Cells and / or STRO-1 expressing TNAP ＋ Any of claims 1 to 17, derived from cells 1. The method according to claim 1.

19. STRO-1 ＋ cells and / or their progeny and / or soluble Claims for treating or delaying the progression of pancreatic dysfunction, where the agent is administered after diagnosis of the disorder. Item 19. The method according to any one of items 1 to 18.

20. Onset and / or progression of pancreatic dysfunction and / or blood glucose levels and / or blood Serum insulin level and / or beta cell number and / or alpha cell number and and / or the number of pancreatic islets and / or the number of PDX-1 expressing cells and / or PDX-1 further comprising monitoring or detecting the amount of expression and / or the number of blood vessels; 20. The method according to any one of claims 1 to 19.

21. STRO-1 ＋ cells and / or their progeny and / or soluble The sex factor is STRO-1 ＋ cells and / or their progeny and / or and administered in the form of a composition comprising the soluble factor derived therefrom and a carrier and / or excipient.

21. The method according to any one of claims 1 to 20.

22. The composition further comprises a factor that induces or enhances differentiation of the progenitor cells into vascular cells, or 22. The method of claim 21, wherein the composition comprises tissue-specific unipotent cells.

23. (i) treatment of pancreatic dysfunction; and / or (ii) improving pancreatic function; and / or (iii) inducing or promoting the regeneration of pancreatic beta cells and / or pancreatic islets; and / or teeth (iv) a decrease in blood glucose levels and / or an increase in blood / serum insulin levels; and / or or (v) increasing the number of pancreatic beta cells and / or increasing the number of pancreatic beta cells relative to pancreatic alpha cells Increased number of cells and / or decreased number of pancreatic alpha cells and / or increased number of pancreatic islets addition; and / or (vi) increased expression of pancreatic duodenal homeobox factor-1 (PDX-1) and / or an increase in the number of PDX-1 expressing cells in the pancreas; and / or (vii) Induction or promotion of arteriogenesis or angiogenesis in the pancreas For STRO-1 ＋ cells and / or their progeny and / or derived therefrom Soluble factors or compositions containing them.

24. (i) treatment of pancreatic dysfunction; and / or (ii) improving pancreatic function; and / or (iii) inducing or promoting the regeneration of pancreatic beta cells and / or pancreatic islets; and / or teeth (iv) a decrease in blood glucose levels and / or an increase in blood / serum insulin levels; and / or or (v) increasing the number of pancreatic beta cells and / or increasing the number of pancreatic beta cells relative to pancreatic alpha cells Increased number of cells and / or decreased number of pancreatic alpha cells and / or increased number of pancreatic islets addition; and / or (vi) increased expression of pancreatic duodenal homeobox factor-1 (PDX-1) and / or an increase in the number of PDX-1 expressing cells in the pancreas; and / or (vii) Induction or promotion of arteriogenesis or angiogenesis in the pancreas For STRO-1 ＋ cells and / or their progeny and / or derived therefrom and the use of soluble factors or compositions containing them.

25. (i) treatment of pancreatic dysfunction; and / or (ii) improving pancreatic function; and / or (iii) inducing or promoting the regeneration of pancreatic beta cells and / or pancreatic islets; and / or teeth (iv) a decrease in blood glucose levels and / or an increase in blood / serum insulin levels; and / or or (v) increasing the number of pancreatic beta cells and / or increasing the number of pancreatic beta cells relative to pancreatic alpha cells Increased number of cells and / or decreased number of pancreatic alpha cells and / or increased number of pancreatic islets addition; and / or (vi) increased expression of pancreatic duodenal homeobox factor-1 (PDX-1) and / or an increase in the number of PDX-1 expressing cells in the pancreas; and / or (vii) Induction or promotion of arteriogenesis or angiogenesis in the pancreas For STRO-1 ＋ cells and / or their progeny and / or derived therefrom Use of a soluble factor in the manufacture of a pharmaceutical product.

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