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Efficient pre-treatment device for culturing human amniotic mesenchymal stem cells and culture method thereof

A pretreatment device and stem cell technology, applied in the field of human cell culture, can solve the problems of inability to be popularized and applied on a large scale, high consumption of manpower and material resources, low efficiency, etc. low cost effect

Active Publication Date: 2017-12-15
上海莱威生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when human amniotic membrane tissue is pretreated, human amniotic membrane tissue needs to be pretreated in different processing equipment or containers, and human amniotic membrane tissue is easily polluted by microorganisms such as bacteria and viruses in the air in this large space transfer overshoot Therefore, the current culture of human amniotic zone mesenchymal stem cells under this open pretreatment method is cumbersome, inefficient, consumes a lot of manpower and material resources, and has a high growth rate, so it cannot be popularized and applied on a large scale.

Method used

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  • Efficient pre-treatment device for culturing human amniotic mesenchymal stem cells and culture method thereof
  • Efficient pre-treatment device for culturing human amniotic mesenchymal stem cells and culture method thereof
  • Efficient pre-treatment device for culturing human amniotic mesenchymal stem cells and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1 as figure 1 As shown, this embodiment provides a high-efficiency pretreatment device for culturing human amniotic mesenchymal stem cells, which specifically includes: an upper cover 100 that can be turned over on a horizontal axis, and a grid-shaped knife groove is opened on the upper cover 100 101; the casing 200 hingedly connected with the upper cover 100, the casing 200 is square or circular, the upper cover 100 is arranged in axial connection with the top rear side of the casing 200; the cutting assembly 300 located at the opening of the casing 200, the cutting assembly 300 Set in cooperation with the upper cover 100, through the pressing force of the upper cover 100 on the cutting assembly 300, the washed human amniotic membrane is evenly placed on the cutting assembly 300 and cut into small tissue pieces of the same size; located in the middle of the housing 200 Digestion assembly 400, that is, the digestion assembly 400 is located below the cutting asse...

Embodiment 2

[0077] Example 2 This example provides a method for pretreatment of human amniotic mesenchymal stem cells using the high-efficiency pretreatment device of Example 1, including the following steps:

[0078] Step 1) Acquisition of human amniotic membrane tissue

[0079] To obtain neonatal amniotic membrane tissue, try to use cesarean section newborn amniotic membrane to avoid contamination, put human amniotic membrane in ice saline, keep it at a low temperature of about 4 degrees, and quickly bring it back to the cell laboratory. In the cabinet, the amniotic membrane slices were fully washed with iced saline to remove blood and impurities, and then the amniotic tissue was repeatedly folded into 3-5 layers of 5cm x 5cm tissue blocks, and the tissue blocks were placed in aseptically treated On the cutting assembly 300, select the cutting specifications of the grid-shaped blade 301 and the knife groove 101 according to the needs, press the upper cover 100 to cut the tissue block in...

Embodiment 3

[0084] Example 3 This example provides a method for culturing human amniotic mesenchymal stem cells pretreated in Example 2 above, including the following steps:

[0085] Step 1) Culture medium preparation

[0086] Prepare DMEM / F12 medium containing 20% ​​fetal bovine serum and 1% penicillin-streptomycin, and DMEM / F12 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin respectively; On the first day of primary culture, subculture and recovery, use DMEM / F12 medium containing 20% ​​fetal bovine serum and 1% penicillin and streptomycin; at other times, use 10% fetal bovine Serum, 1% penicillin and streptomycin in DMEM / F12 medium for culture;

[0087] Step 2) Two-dimensional culture

[0088] Take the relatively pure human amniotic mesenchymal stem cells pretreated in Example 2, and resuspend the relatively pure human amniotic mesenchymal stem cells in the above-mentioned medium to prepare amniotic mesenchymal stem cell resuspension, which is added to T175 cu...

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Abstract

The invention discloses an efficient pre-treatment device for culturing human amniotic mesenchymal stem cells and a culture method thereof. The efficient pre-treatment device comprises an upper cover, a shell, a cutting assembly, a digestion assembly and a cleaning centrifugal part, wherein the shell is hinged to the upper cover; the cutting assembly located at an opening of the shell is matched with the upper cover; the digestion assembly is located in the middle of the shell; and the cleaning centrifugal part is located on the lower portion of the shell and comprises a U-shaped cabin body, a stirring paddle and a micromotor. The invention also provides a cell pre-treatment and culture method based on the efficient pre-treatment device. The method comprises the following six steps: human amniotic membrane cutting and separating; enzymic digestion; centrifugal purification; culture medium preparation; two-dimensional culture; and subculture. The cell culture method based on the efficient pre-treatment device disclosed by the invention is stable in technology and good in repeatability, avoids the problems of low cell activity and poor yield, improves the cell activity and yield greatly, simplifies the operating process, and is flexible and convenient to use.

Description

technical field [0001] The invention relates to the technical field of human cell culture, in particular to a high-efficiency pretreatment device and a culture method for culturing human amniotic mesenchymal stem cells. Background technique [0002] Pluripotent mesenchymal cells, also known as mesenchymal stem cells, are a kind of very characteristic adult stem cells. These cells have the potential to differentiate into various mature cells, including: adipocytes, chondrocytes, bone cells, tenocytes and muscle cells. These properties, as well as their developmental plasticity, have attracted widespread attention in the scientific community, as they could be used in the field of regenerative medicine to replace degenerated and dead tissues. Mesenchymal stem cells can differentiate into chondrocytes in serum-free culture with transforming growth factor, and osteoblasts in serum with ascorbic acid and dexamethasone. Mesenchymal stem cells have the property of proliferating ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/40C12M1/02C12M1/00C12N5/0775
CPCC12M21/18C12M27/02C12M45/02C12N5/0668C12N2509/00
Inventor 王晓冰
Owner 上海莱威生物科技有限公司
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