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Use of PLGA material in building tissue engineering liver cells based on ADSCs

A tissue engineering and basic technology, applied in the field of PLGA materials, can solve the problems of late research and slow progress of ADSCs, and achieve the effect of broad application prospects.

Inactive Publication Date: 2012-10-03
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the late and slow progress in the field of ADSCs differentiation into hepatocytes, there is no report on the differentiation of ADSCs into hepatocytes on any three-dimensional scaffold.

Method used

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  • Use of PLGA material in building tissue engineering liver cells based on ADSCs
  • Use of PLGA material in building tissue engineering liver cells based on ADSCs
  • Use of PLGA material in building tissue engineering liver cells based on ADSCs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Inducing ADSCs to differentiate into hepatocytes on the surface of the PLGA scaffold in vitro

[0024] Using the method of the present invention to induce ADSCs (taking hADSCs as an example) to differentiate into hepatocytes in vitro comprises the following steps:

[0025] 1. Isolation and culture of hADSCs

[0026] ① Take the fat sucked out and discarded during liposuction, and wash it repeatedly with an equal volume of PBS (pH7.0-7.2) buffer to remove blood cells and anesthetics.

[0027] ②Digest with PBS (pH7.0-7.2) containing 0.075% (V / V) collagenase at 37°C for 40min, and gently shake during digestion to release intercellular connections.

[0028] ③ The collagenase was neutralized with an equal volume of low-sugar DMEM (purchased from Gibco) containing 10% (V / V) fetal bovine serum (purchased from HyClone).

[0029] ④Centrifuge at 1500rpm for 5min after filtering with a 200-mesh sieve, and discard the supernatant.

[0030] ⑤ Dilute the cell pellet with...

Embodiment 2

[0038] Example 2, identification of differentiated hepatocytes

[0039] The cells induced to differentiate in Example 1 were identified by the following methods:

[0040] 1. DAPI staining

[0041] In order to evaluate the distribution of cells throughout the material, the PLGA scaffold seeded with hADSCs in Example 1 and induced with hepatocyte induction medium for 5 days was prepared into frozen sections, and then stained with DNA-specific fluorescent dye DAPI. The DAPI staining results of hADSCs induced on the surface of PLGA scaffolds for 5 days were as follows: figure 1 As shown (20×), hADSCs were evenly distributed in the PLGA scaffold and integrated with the scaffold, indicating that the PLGA scaffold can well support the adhesion of hADSCs in vitro.

[0042] 2. Detection by CCK8 method

[0043] The CCK8 method was used to evaluate the cell growth of hADSCs in the induction process of hepatocytes. The method was: in Example 1, 10 μl CCK8 was added to 100 μl culture me...

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Abstract

The invention discloses an application of a PLGA material in the origination project liver unit construction based on ADSCs. The application is that the ADSCs are inoculated to the surface of the PLGA material for carrying out induction on liver cells. Results of morphological observations, gene and protein levels and function detections and the like prove that the PLGA stent supports the adhesion and proliferation of the ADSCs and the differentiation to the liver cells. The invention provides a stent material used for the origination project liver unit construction based on the ADSCs, which has wide application prospect.

Description

technical field [0001] The invention relates to a new application of the PLGA material, that is, it can be used as a scaffold material for constructing tissue engineering liver units based on ADSCs. Background technique [0002] Similar to bone marrow, there are pluripotent stem cells in adipose tissue, called adipose-derived stem cells (ADSCs). ADSCs can differentiate into various cells such as chondrocytes, osteoblasts, adipocytes, cardiomyocytes and nerve cells, and are widely used in tissue engineering. Compared with other stem cells, ADSCs have many advantages as a cell source for tissue engineering. For example, during autologous treatment, ADSCs obtained from patients are less invasive, and autologous ADSCs do not have the problem of transplant rejection. In addition, because ADSCs are easy to obtain in large quantities and expand rapidly in vitro, it is easy to reach the number of cells required for clinical practice. [0003] Recently, it has been reported that AD...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/18A61F2/02
Inventor 裴雪涛王敏裴海云师伟管利东
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY