Stem-cell biological preparation, and preparation method and application thereof

A technology of biological preparations and stem cells, applied in the field of biomedicine, can solve problems such as unsatisfactory treatment effects, achieve the effects of promoting tissue regeneration, improving liver damage, and good therapeutic effects

Active Publication Date: 2015-11-18
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These chemosynthetic drugs can reduce the degenerated necrotic cells at the site of liver injury and shrink the area of ​​liver necrosis, thereby playing a certain therapeutic role in liver injury, but the therapeutic effect is not ideal

Method used

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  • Stem-cell biological preparation, and preparation method and application thereof
  • Stem-cell biological preparation, and preparation method and application thereof
  • Stem-cell biological preparation, and preparation method and application thereof

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preparation example Construction

[0060] The invention provides a method for preparing a stem cell biological preparation, comprising the following steps:

[0061] a) Liver stem cells, peripheral blood stem cells, albumin and a solvent are mixed to obtain a stem cell biological preparation.

[0062] In the preparation method provided by the present invention, stem cell biological preparations can be obtained by directly mixing hepatic stem cells, peripheral blood stem cells, albumin and a solvent, and the process is specifically as follows:

[0063] a1), mixing albumin with a solvent to obtain an albumin solution;

[0064]a2), liver stem cells, peripheral blood stem cells and the albumin solution are mixed to obtain a stem cell biological preparation.

[0065] In the above preparation method provided by the present invention, albumin and a solvent are first mixed to obtain an albumin solution. Wherein, the solvent is preferably physiological saline.

[0066] After the albumin solution is prepared, the album...

Embodiment 1

[0088] Obtain liver stem cells

[0089] The liver tissue was washed several times with PBS, chopped and digested with trypsin solution (trypsin content: 0.25g / 100mL), overnight at 4°C, and taken out the next day at 37°C for 5-10 minutes of further digestion. After digestion, suck out the cell suspension, add type II collagenase solution (type II collagenase content is 0.25g / 100mL) to the remaining tissue, mix well, seal, and digest at 37°C for 10min. The digested tissue was filtered with a 200-mesh sieve, and the filtered cell suspension was centrifuged at 270 g for 5 min. After centrifugation, discard the upper liquid, and wash the lower cell pellet with PBS twice.

[0090] Use the DMEM high-glucose medium (Gibco company, lot number 12100046.) containing 15% (volume percent concentration) FBS to resuspend the cell pellet obtained above, with 1 × 10 6 cells / mL were inoculated into culture flasks. Gently shake the culture flask back and forth to distribute the cell suspensio...

Embodiment 2

[0094] Obtain peripheral blood stem cells

[0095] 1) Separation of peripheral blood mononuclear cells:

[0096] Peripheral blood and normal saline were mixed at a volume ratio of 1:1, and the mixed solution was pipetted, and then centrifuged with Ficoll separation solution (Fresenius Company, batch number: 12BS15) with a density of 1.073 g / mL. The centrifugal force of separation is 700g, and the time of centrifugal separation is 20min. After centrifugation, the buffy coat cells in the middle of the centrifugation product were taken, which were peripheral blood mononuclear cells (MNCs).

[0097] 2) Culture and expansion of peripheral blood stem cells:

[0098] After the peripheral blood mononuclear cells obtained above were counted, they were mixed with DMEM high-glucose medium (Gibco company, batch number: 12100046) containing 10% (volume percentage) FBS to obtain a cell density of 3 × 10 5 cells / mL of cell suspension, inoculate 10 mL of cell suspension into a 10 cm plate,...

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Abstract

The invention belongs to the field of biological medicine and particularly relates to a stem-cell biological preparation, and a preparation method and an application thereof. The stem-cell biological preparation comprises liver stem cells, peripheral blood stem cells, albumin and a solvent. The stem-cell biological preparation contains the liver stem cells, the peripheral blood stem cells and the albumin, and by the synergistic effect of the liver stem cells, the peripheral blood stem cells and the albumin, the tissue regeneration of the injured part of the liver can be effectively promoted, so that better liver-injury treatment effect is realized. Shown by experimental results, when the stem-cell biological preparation provided by the invention is adopted for treating a rat with acute liver injury, ALT, AST and TBIL indexes in the blood of the rat are respectively and obviously reduced, so that the liver injury of the rat can be obviously improved by adopting the biological preparation provided by the invention.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a stem cell biological preparation and its preparation method and application. Background technique [0002] The liver is an organ in the body that mainly has metabolic functions, and plays a role in the body in deoxidation, storage of glycogen, synthesis of secretory proteins, and so on. The liver also produces bile in the digestive system. In medical terms, the liver is often described with the Latin prefix Hepato- or Hepatic. The liver is the largest organ in the human body. It is located in the abdomen in the human body, under the right diaphragm, in front of the gallbladder, in front of the right kidney, and above the stomach. Due to its large size and brittle texture, the liver is extremely vulnerable to damage. [0003] Liver injury is a common disease all over the world. Severe liver injury will develop into liver cirrhosis and liver cancer, which seriously threa...

Claims

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Application Information

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IPC IPC(8): C12N5/074C12N5/0789A61K35/12A61K38/38A61P1/16
Inventor 陈海佳王一飞葛啸虎卢瑞珊李平
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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