Preparation method of deciduas mesenchymal stem cells

A technology of stem cells and cells, applied in the field of cell biology, can solve problems such as difficulty in realizing large-scale production, and achieve the effects of strong immune regulation, short time consumption and low cost.

Inactive Publication Date: 2018-07-06
重庆斯德姆生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the actual production is limited by factors such as t

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  • Preparation method of deciduas mesenchymal stem cells
  • Preparation method of deciduas mesenchymal stem cells
  • Preparation method of deciduas mesenchymal stem cells

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Embodiment 1

[0037] This embodiment provides a method for preparing decidual mesenchymal stem cells, which specifically includes the following steps:

[0038] Step 1: Separation: After washing the placental tissue with sterile saline, carefully separate the umbilical cord and placental decidua tissue, and separate the umbilical cord and decidua tissue;

[0039] Step 2: Initial treatment: Cut the umbilical cord and placental decidua tissues in step 1 into small tissue pieces, wash away the blood components in the two, and then add type II collagenase and trypsin to digest the small tissue pieces.

[0040] Step 3: Filtration: Filter the mixed liquid obtained after digestion through a 200-mesh filter to remove insoluble substances.

[0041] Step 4: Inoculation and cultivation: inoculate the filtered mixed liquid into cell culture flasks respectively, and cultivate them in an incubator at 37° C., 5% CO 2 , and saturated humidity with DF12 culture liquid containing fetal bovine serum.

[0042] S...

Embodiment 2

[0046] This embodiment provides a method for preparing decidual mesenchymal stem cells, which specifically includes the following steps:

[0047] Step 1: Separation: After washing the placental tissue with sterile saline, carefully separate the umbilical cord and placental decidua tissue, and separate the umbilical cord and decidua tissue;

[0048] Step 2: Initial treatment: Cut the umbilical cord and placental decidua tissues in step 1 into small tissue pieces, wash away the blood components in the two, and then add type I collagenase and trypsin to digest the small tissue pieces.

[0049] Step 3: Filtration: Filter the mixed liquid obtained after digestion through a 200-mesh filter to remove insoluble substances.

[0050] Step 4: Inoculation and cultivation: Inoculate the filtered mixed liquid into cell culture flasks respectively, and use DF12 culture medium containing fetal bovine serum at 37°C and 6% CO 2 cultured in a humidified incubator.

[0051] Step 5: Subculture: ...

Embodiment 3

[0055] This embodiment provides a method for preparing decidual mesenchymal stem cells, which specifically includes the following steps:

[0056] Step 1: Separation: After washing the placental tissue with sterile saline, carefully separate the umbilical cord and placental decidua tissue, and separate the umbilical cord and decidua tissue;

[0057] Step 2: Initial treatment: Cut the umbilical cord and placental decidua tissues in step 1 into small tissue pieces, wash away the blood components in the two, and then add type II collagenase and trypsin to digest the small tissue pieces.

[0058] Step 3: Filtration: Filter the mixed liquid obtained after digestion through a 200-mesh filter to remove insoluble substances.

[0059] Step 4: inoculation and cultivation: Inoculate the filtered mixed liquid into cell culture flasks respectively, and cultivate them in an incubator at 37° C., 4% CO 2 , and saturated humidity with DF12 culture liquid containing fetal bovine serum.

[0060]...

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Abstract

The invention discloses a preparation method of deciduas mesenchymal stem cells. The preparation method comprises the following steps: step 1, separation; step 2, initial treatment; step 3, filtration; step 4, inoculation and culture; step 5, passage; step 6, cell cryopreservation; step 7, cell thawing. According to the preparation method, the entire process from the start of raw material collection to completion of cell preparation is concentratedly and systematically monitored, and the quality from the start of screening before placenta collection to cellpreparation of a finished intermediate product and a final product is all controlled.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for preparing mesenchymal stem cells. Background technique [0002] Stem cells are a kind of pluripotent cells with self-replication ability. Under certain conditions, it can differentiate into a variety of functional cells. Stem cells are divided into embryonic stem cells (embryonic stem cells, ES cells) and adult stem cells (somatic stem cells) according to their developmental stages. Stem cells are divided into three categories according to their developmental potential: totipotent stem cells (TSCs), pluripotent stem cells (pluripotent stem cells), and unipotent stem cells (unipotent stem cells). Stem cells are not fully differentiated and immature cells, which have the potential to regenerate various tissues and organs and the human body. They are called "universal cells" in the medical field. Stem cells exist in all multicellular tissues, can divide into a variety of ...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A01N1/02
CPCC12N5/0668A01N1/0221C12N2500/90C12N2509/00
Inventor 宋彩霞熊华强杨仁淑秦连荣
Owner 重庆斯德姆生物技术有限公司
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