Human pluripotent stem cell-derived islet cell preparation and preparation method thereof

Abstract

The human pluripotent stem cell-derived islet cell preparation provided by the invention can maintain the structure and biological activity of human pluripotent stem cell-derived islet cells in the transportation process, and can be used for preparing injections, and the finished product meets the injection requirements of sterility, no pyrogen, safety, stability and the like.

Description

Technical Field

[0001] The present invention relates to the field of stem cells. Specifically, the present invention provides a preparation of islet cells derived from human pluripotent stem cells and a method for preparing the same, which can be used to treat insulin deficiency-related diseases by injection through the hepatic portal vein or under the rectus sheath. Background Art

[0002] β-cell (BC) transplantation may offer a definitive cure for type 1 diabetes. However, the limited availability of donor β-cells restricts the use of this treatment as a clinical therapy. Pluripotent (PS) cells can proliferate indefinitely and differentiate into multiple cell types. Therefore, PS cells are a promising source of β-cells. Islet cells derived from human pluripotent stem cells are cell spheres with a cell diameter greater than 50 μm obtained by directing the differentiation of human pluripotent stem cells. Currently, there is no report on formulating islet cells derived from human pluripotent stem cells into a preparation. After the directed differentiation of stem cell-derived islet cells is completed in a cell factory, they need to be transported to a medical institution for transplantation of islet cells derived from human pluripotent stem cells, and the problem of decreased cell viability will be faced during transportation. Summary of the Invention

[0003] The preparation of islet cells derived from human pluripotent stem cells provided by the present invention is easy to transport over long distances, can maintain the structure and biological activity of islet cells derived from human pluripotent stem cells during long-distance transportation, and can be used to prepare an injection; it is particularly suitable for treating insulin deficiency-related diseases by injection through the hepatic portal vein or under the rectus sheath after long-distance transportation. After the preparation is formulated into an injection, the finished product meets the requirements of injections such as sterility, pyrogen-free, safety, and stability.

[0004] By providing a preparation of islet cells derived from human pluripotent stem cells, the present invention can maintain the ratio of NKX6.1+ and C-PEP+ double-positive cells in the islet cells derived from human pluripotent stem cells in the preparation and / or maintain a high level of biological activity.

[0005] In a first aspect, the present invention provides a preparation of islet cells derived from human pluripotent stem cells.

[0006] In some embodiments, the preparation of islet cells derived from human pluripotent stem cells comprises islet cells derived from pluripotent stem cells (hPSC), human serum albumin, and a basal solvent, wherein the mass concentration of human serum albumin is 0.625%-5%. The applicant has found that when the concentration of added human serum albumin is lower than 0.625%, the biological activity of the stored islet cells derived from human pluripotent stem cells is lower than 90%.

[0007] In some embodiments, the base solvent in the human pluripotent stem cell-derived islet cell preparation is selected from 0.9% normal saline, sodium lactate Ringer's solution (lactated Ringer's solution), 5% glucose, or a mixed solvent thereof. The inventors have found through research that when the base solvent is selected from phosphate buffer solution, the cell morphology and activity are significantly reduced.

[0008] In some embodiments, in each milliliter of the human pluripotent stem cell-derived islet cell preparation, the content of human pluripotent stem cell-derived islet cells is 10 million to 15 million single cells, preferably 4 million to 12 million single cells. The applicant has found that when the content of human pluripotent stem cell-derived islet cells is at a concentration of 1 million single cells, the cell activity and morphology are poor.

[0009] In some embodiments, in each milliliter of the human pluripotent stem cell-derived islet cell preparation, the content of human pluripotent stem cell-derived islet cells is 1000 to 10000 IEQ, preferably 2000 to 8000 IEQ, more preferably 2500 to 7000 IEQ. The applicant has found that when the content of human pluripotent stem cell-derived islet cells is less than 1000 IEQ, the cell activity and morphology are poor.

[0010] Note: One IEQ of human pluripotent stem cell-derived islet cells is equivalent to 1000 - 2000 single cells.

[0011] In some embodiments, the human pluripotent stem cell-derived islet cell preparation further comprises a stabilizer, and the stabilizer is selected from one or several of HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), pyruvate, nicotinamide, glucose, zinc sulfate, ITS (1X), linoleic acid, vitamin E. Preferably, it is selected from one or several of HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid), pyruvate, nicotinamide, glucose. More preferably, the following combinations:

[0012] i. The combination of 25 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 7.5 mM pyruvate, 10 mM nicotinamide, 4.5 mM glucose; or

[0013] ii. The combination of 25 mM HEPES, 7.5 mM sodium pyruvate, 10 mM nicotinamide; or iii. The combination of 7.5 mM sodium pyruvate, 10 mM nicotinamide, 4.5 mM glucose; or iv. The combination of 25 mM HEPES, 7.5 mM sodium pyruvate, 4.5 mM glucose; or

[0014] v. The combination of 25 mM HEPES, 5 mM sodium pyruvate, 0.2 - 10 mM nicotinamide, 16.7 mM zinc sulfate, 1X ITS; or

[0015] vi. A composition of 25 mM HEPES, 5 mM sodium pyruvate, 10 mM nicotinamide, 16.7 mM zinc sulfate, 1X ITS, 0.019 μM linoleic acid, and 10 μM vitamin E.

[0016] In some embodiments, the human pluripotent stem cell-derived islet cell preparation further comprises a pH regulator to control the pH of the preparation within the range of 6.5 - 8.0.

[0017] In some embodiments, the human pluripotent stem cell-derived islet cells in the human pluripotent stem cell-derived islet cell preparation are double positive for NKX6.1+ and C-PEP+;

[0018] Preferably, after the preparation is stored at 16°C to 22°C (±2°C) for 24 hours, at least 20% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+; preferably, at least 30% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, at least 35% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, at least 40% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, at least 45% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, at least 50% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, and at least 55% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+. The specific data can be selected from any integer or decimal value between at least 20% and 95%.

[0019] In some embodiments, the pluripotent stem cell-derived islet cells in the human pluripotent stem cell-derived islet cell preparation are cell spheres larger than 50 μm; preferably cell spheres of 50 - 350 μm. The specific values are selected from any integer or decimal value within 50 μm - 350 μm, such as 50 μm, 100 μm, 150 μm, 200 μm, 250 μm, 300 μm, 350 μm.

[0020] In some embodiments, for the human pluripotent stem cell-derived islet cell preparation, when stored at 16°C to 22°C (±2°C) for 24 hours, the human pluripotent stem cell-derived islet cells maintain more than 85% biological activity; preferably maintain more than 90%, 92.5%, 95% biological activity. The specific values are selected from any integer or decimal value between 85% and 95%.

[0021] In some embodiments, for the human pluripotent stem cell-derived islet cell preparation, when stored at 16°C to 22°C (±2°C) for 24 hours, it can maintain good cell morphology. The good cell morphology is manifested as a smooth cell sphere surface, uniform size, and good refraction.

[0022] In some embodiments, for a preparation of human pluripotent stem cell-derived islet cells, when stored at 16°C to 22°C (±2°C) for 24 hours, the human pluripotent stem cell-derived islet cells can maintain a good proportion of NKX6.1+ and C-PEP+ double-positive cells, biological activity, and cell morphology.

[0023] In some embodiments, the preparation of human pluripotent stem cell-derived islet cells is an injection. In some embodiments, the injection is preferably for portal vein injection or injection into the anterior sheath of the rectus abdominis muscle, more preferably an injection preparation under the anterior sheath of the rectus abdominis muscle.

[0024] On the other hand, the present invention provides a method for preparing the preparation of human pluripotent stem cell-derived islet cells according to any one of the foregoing embodiments, comprising the following steps:

[0025] (1) Preparation of cell preservation composition: Add human albumin to a basic solvent and mix evenly to prepare a preservation composition for human pluripotent stem cell-derived islet cells.

[0026] (2) Mix the cell preservation composition with human pluripotent stem cell-derived islet cells and fill into a packaging container.

[0027] On the other hand, the present invention provides the use of the preparation of human pluripotent stem cell-derived islet cells according to any one of the foregoing embodiments in the preparation of a drug for treating insulin deficiency. In some embodiments, preferably the drug is a portal vein injection or an injection preparation under the rectus abdominis sheath, more preferably an injection preparation under the anterior rectus abdominis sheath.

[0028] On the other hand, the present invention provides the use of a preservation composition for human pluripotent stem cell-derived islet cells in long-distance transportation, wherein the preservation composition is the component other than human pluripotent stem cell-derived islet cells in the preparation according to any one of the foregoing embodiments.

[0029] In some embodiments, the long-distance transportation temperature is 4°C to 37°C; the transportation time does not exceed 96 hours. In some embodiments, the long-distance transportation temperature is preferably 4°C, 10°C, 16°C, 22°C, 28°C, 37°C. In some embodiments, the transportation time is preferably not more than 96 hours, 48 hours, 36 hours, 24 hours, 20 hours, 16 hours, 12 hours, 8 hours, 6 hours, or 4 hours.

[0030] For the differentiation and preparation methods of human pluripotent stem cell-derived islet cells in the present invention, refer to WO2023 / 097513A1 and PCT / CN2023 / 096328. The entire contents of WO2023 / 097513A1 and PCT / CN2023 / 096328 are incorporated herein by reference, especially regarding the differentiation and preparation methods of human pluripotent stem cell-derived islet cells. Detailed implementation manners

[0031] The solutions of the present disclosure will be explained below in conjunction with embodiments. Those skilled in the art will understand that the following embodiments are only used to illustrate the present disclosure and should not be construed as limiting the scope of the present disclosure. For those embodiments where specific technologies or conditions are not indicated, the technologies or conditions described in the literature in the art or according to the product specifications are followed. For reagents or instruments whose manufacturers are not indicated, they are all conventional products that can be obtained through commercial purchases.

[0032] Detection method

[0033] After 0 - 96 hours of the preparation, the morphology and size of human pluripotent stem cell-derived islet cells were observed under a microscope, and the diameter size distribution was analyzed using CellView.

[0034] Through NC-200 TM The cell viability was detected by a cell counter. Specifically: the human pluripotent stem cell-derived islet cells were digested into single cells using Accutase, and the Via1-Cassette TM was inserted into the device card slot, and NC-200 TM was used for detection with a cell counter.

[0035] The expression of markers related to the biological activity of human pluripotent stem cell-derived islet cells was detected by flow cytometry, such as the proportion of β cells (NKX6.1 & c-pep), the proportion of endocrine cells (CHGA), and the proportion of α cells (ARX & GCG). Specifically: the human pluripotent stem cell-derived islet cells were digested into single cells using Accutase, fixed overnight with BD Cytofix / Cytoperm TM solution, permeabilized with Wash Buffer, and then incubated with antibodies. After washing, it was filtered through a cell sieve and detected with a flow cytometer.

[0036] Example 1

[0037] The mass concentration of human serum albumin in the human pluripotent stem cell-derived islet cell preparation is 1%, the basic solvent is physiological saline, and the concentration of physiological saline is 0.9%. The content of human pluripotent stem cell-derived islet cells in each milliliter of the human pluripotent stem cell-derived islet cell preparation is 1×10 7 cells. The human pluripotent stem cell-derived islet cells used in the embodiments of the present invention were all prepared by the method described on pages 36 - 37, Stage 1 - 6 of WO2023 / 097513A, and the diameter of the cell spheres is 50 - 350 μm.

[0038] Preparation method

[0039] 1. Preparation of cell preservation composition: Using 0.9% normal saline as the base solvent, add human albumin, and mix evenly to prepare a human pluripotent stem cell-derived islet cell preservation composition.

[0040] 2. Prepare a preparation by mixing the cell preservation composition and human pluripotent stem cell-derived islet cells at a concentration of 1*10 7 cells / ml, fill it into a preparation bag, and store it at 22°C (±2°C).

[0041] Cell viability assay

[0042] 1. Verification: Take the finished product prepared in Example 1, store it in an incubator at 22°C (±2°C), and take out the human pluripotent stem cell-derived islet cells for detection after 24 hours of storage.

[0043] 2. Detection items: Cell morphology, cell surface markers, and cell viability. The test results are summarized in Table 1.

[0044] Table 1 Test results of the preparation in Example 1

[0045]

[0046] Example 2

[0047] This example provides a human pluripotent stem cell-derived islet cell preparation, which includes human pluripotent stem cell-derived islet cells, human albumin, a base solvent, and a stabilizer. The mass concentration of human albumin in the human pluripotent stem cell-derived islet cell preparation is 1%, the base solvent is normal saline, and the concentration of normal saline is 0.9%. The content of human pluripotent stem cell-derived islet cells in each milliliter of the human pluripotent stem cell-derived islet cell preparation is 1*10 7 cells. The stabilizer includes a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid solution with a molar concentration of 25 mM, a sodium pyruvate solution with a molar concentration of 7.5 mM, a nicotinamide solution with a molar concentration of 10 mM, and a glucose solution with a molar concentration of 4.5 mM.

[0048] Preparation method

[0049] 1. Preparation of cell preservation composition: Using 0.9% normal saline as the base solvent, add human albumin, then add the stabilizer, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, sodium pyruvate, nicotinamide, and glucose, mix evenly, and then sterilize and filter using a 220 nm filter to prepare a human pluripotent stem cell-derived islet cell preservation composition.

[0050] 2. Prepare a preparation by mixing the cell preservation composition and human pluripotent stem cell-derived islet cells at a concentration of 1*10 7Prepare a preparation at a concentration of cells / ml, fill it into a preparation bag, and store it at 22°C (±2°C).

[0051] Cell viability assay

[0052] 1. Assay: Take the finished product prepared in Example 2, store it in an incubator at 22°C (±2°C), and take out the human pluripotent stem cell-derived islet cells for detection after 24 hours of storage.

[0053] 2. Detection items: cell morphology, cell surface markers, and cell viability.

[0054] Example 3

[0055] Compared with Example 2, the only difference is the concentration of human albumin. The concentration of human albumin in the human pluripotent stem cell-derived islet cell preparation in this example is 0.625%, and the other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0056] Example 4

[0057] Compared with Example 2, the only difference is the concentration of human albumin. The concentration of human albumin in the human pluripotent stem cell-derived islet cell preparation in this example is 2.50%, and the other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0058] Example 5

[0059] Compared with Example 2, the only difference is the concentration of human albumin. The concentration of human albumin in the human pluripotent stem cell-derived islet cell preparation in this example is 5%, and the other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0060] Example 6

[0061] Compared with Example 2, the only difference is that the content of human pluripotent stem cell-derived islet cells is 6*10 6 cells / ml. The other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0062] Example 7

[0063] Compared with Example 2, the only difference is that the content of human pluripotent stem cell-derived islet cells is 8*10 6 cells / ml. The other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0064] Example 8

[0065] Compared with Example 2, the only difference is that the storage temperature of the preparation is 16°C (±2°C), and the other components in the islet cell preparation are the same as those in Example 2, the preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0066] Comparative Example 1

[0067] Compared with Example 2, the only difference is that in this comparative example, the stabilizer only adds a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid solution with a molar concentration of 25 mM, the preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0068] Comparative Example 2

[0069] Compared with Example 2, the only difference is that in this comparative example, the stabilizer adds a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid with a molar concentration of 25 mM and a glucose solution with a concentration of 4.5 mM, the preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0070] Comparative Example 3

[0071] Compared with Example 2, the only difference is that in this comparative example, the stabilizer adds a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid with a molar concentration of 25 mM and a sodium pyruvate solution with a concentration of 7.5 mM, the preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0072] Comparative Example 4

[0073] Compared with Example 2, the only difference is that in this comparative example, the stabilizer adds a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid with a molar concentration of 25 mM, a sodium pyruvate solution with a concentration of 7.5 mM and a nicotinamide solution with a concentration of 10 mM, the preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0074] Table 2 Test results of Example 2 and Comparative Examples 1-4

[0075]

[0076] Comparative Example 5

[0077] Compared with Example 2, the only difference is the concentration of human serum albumin. The human pluripotent stem cell-derived islet cell preparation in this example contains human serum albumin at a concentration of 0.10%, and the other components in the islet cell preparation are the same as those in Example 2, the preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0078] Comparative Example 6

[0079] Compared with Example 2, the only difference is the concentration of human albumin. The concentration of human albumin in the human pluripotent stem cell-derived islet cell preparation in this example is 0.25%, and the other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0080] Table 3 Test results of Examples 2-5 and Comparative Examples 5-6

[0081]

[0082] Comparative Example 7

[0083] Compared with Example 2, the only difference is that the content of human pluripotent stem cell-derived islet cells is 4×10 6 cells / ml. The other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0084] Comparative Example 8

[0085] Compared with Example 2, the only difference is that the content of human pluripotent stem cell-derived islet cells is 2×10 6 cells / ml. The other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0086] Comparative Example 9

[0087] Compared with Example 2, the only difference is that the storage temperature of the preparation is 4°C. The other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0088] Table 4 Test results of Example 2, 6, 7 and Comparative Examples 7-8

[0089]

[0090] Comparative Example 10

[0091] Compared with Example 2, the only difference is that the storage temperature of the preparation is 10°C (±2°C). The other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0092] Comparative Example 11

[0093] Compared with Example 2, the only difference is that the storage temperature of the preparation is 28°C (±2°C). The other components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

[0094] Comparative Example 12

[0095] Compared with Example 2, the only difference is that the storage temperature of the preparation is 37°C (±2°C), and the remaining components in the islet cell preparation are the same as those in Example 2. The preparation method is the same as that in Example 2, and the cell activity experiment is the same as that in Example 2.

Claims

1. A human pluripotent stem cell-derived pancreatic islet cell preparation, comprising pancreatic islet cells derived from pluripotent stem cells (hPSC), human serum albumin and a basic solvent, wherein the mass concentration of human serum albumin is 0.625%-5%.

2. The pancreatic islet cell preparation derived from human pluripotent stem cells as claimed in claim 1, wherein the base solvent is selected from 0.9% saline, sodium lactate Ringer's solution, 5% glucose or a mixed solvent thereof.

3. The human pluripotent stem cell-derived pancreatic islet cell preparation according to claim 1 or 2, wherein the content of human pluripotent stem cell-derived pancreatic islet cells per milliliter of the preparation is 1 to 15 million single cells, preferably 4 to 12 million single cells.

4. The human pluripotent stem cell-derived pancreatic islet cell preparation according to any one of claims 1 to 3, further comprising a stabilizer, wherein the stabilizer is selected from one or more of HEPES (4-hydroxyethylpiperazineethanesulfonic acid), pyruvate, niacinamide, glucose, zinc sulfate, ITS (1X), linoleic acid, and vitamin E; Preferably, one or more of HEPES (4-hydroxyethylpiperazineethanesulfonic acid), pyruvate, niacinamide, and glucose; More preferably, a combination of 25 mM 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), 7.5 mM pyruvate, 10 mM niacinamide, 4.5 mM glucose; or A combination of 25 mM HEPES, 7.5 mM sodium pyruvate, and 10 mM niacinamide; or A combination of 7.5 mM sodium pyruvate, 10 mM niacinamide, and 4.5 mM glucose; or A combination of 25 mM HEPES, 7.5 mM sodium pyruvate, 4.5 mM glucose; or A combination of 25 mM HEPES, 5 mM sodium pyruvate, 0.2-10 mM niacinamide, 16.7 mM zinc sulfate, and 1XITS; or A combination of 25mM HEPES, 5mM sodium pyruvate, 10mM niacinamide, 16.7mM zinc sulfate, 1XITS, 0.019uM linoleic acid, and 10uM vitamin E. 5 . The human pluripotent stem cell-derived pancreatic islet cell preparation according to claim 1 , further comprising a pH adjuster to control the pH of the preparation within the range of 6.5-8.

0.

6. The human pluripotent stem cell-derived pancreatic islet cell preparation according to any one of claims 1 to 6, wherein the human pluripotent stem cell-derived pancreatic islet cells are NKX6.1+ and C-PEP+ double positive; Preferably, after the preparation is stored at 16°C to 22°C (±2°C) for 24 hours, at least 20% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+; preferably, at least 30% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, at least 35% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, at least 40% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, at least 45% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, at least 50% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+, and at least 55% of the human pluripotent stem cell-derived islets are double positive for NKX6.1+ and C-PEP+. 7 . The human pluripotent stem cell-derived pancreatic islet cell preparation according to claim 1 , wherein the pluripotent stem cell-derived pancreatic islet cells are cell spheres larger than 50 um, preferably cell spheres of 50-350 um.

8. The pancreatic islet cell preparation derived from human pluripotent stem cells according to any one of claims 1 to 7, which is stored at 16°C to 22°C (±2°C) for 24 hours, and the pancreatic islet cells derived from human pluripotent stem cells maintain more than 85% of their biological activity; preferably maintain more than 90%, 92.5%, or 95% of their biological activity.

9. The human pluripotent stem cell-derived pancreatic islet cell preparation according to any one of claims 1 to 8, wherein the preparation is an injection, preferably the preparation is a portal vein injection or an anterior rectus sheath injection, more preferably an anterior rectus sheath injection.

10. The method for preparing a pancreatic islet cell preparation derived from human pluripotent stem cells according to any one of claims 1 to 9, comprising the following steps: (1) Preparation of cell preservation composition: adding human serum albumin to a basic solvent and mixing well to prepare a human pluripotent stem cell-derived pancreatic islet cell preservation composition; (2) Mixing the cell preservation composition and pancreatic islet cells derived from human pluripotent stem cells and filling the mixture into a packaging container.

11. Use of the pancreatic islet cell preparation derived from human pluripotent stem cells according to any one of claims 1 to 9 in the preparation of a drug for treating insulin deficiency; Preferably, the drug is a portal vein injection or a subrectus sheath injection preparation, and more preferably, the drug is an anterior subrectus sheath injection preparation.

12. Use of a human pluripotent stem cell-derived pancreatic islet cell preservation composition in long-distance transportation, wherein the preservation composition is the component other than the human pluripotent stem cell-derived pancreatic islet cells in the preparation according to any one of claims 1 to 9, and the long-distance transportation temperature is 4° C. to 37° C.; and the transportation time does not exceed 96 hours; The long-distance transport temperature is preferably 4°C, 10°C, 16°C, 22°C, 28°C, 37°C; The transport time is preferably no more than 96 hours, 48 ​​hours, 36 hours, 24 hours, 20 hours, 16 hours, 12 hours, 8 hours, 6 hours or 4 hours.

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