Method for preparing mesenchymal stem cell

A technique for stromal stem cells and cell culture, applied in the field of mesenchymal stem cell preparation, which can solve the problems of reduced stem cell number, proliferation and differentiation ability, inability to extract bone marrow, and patient injury.

Active Publication Date: 2013-11-20
SHUNHAO CELL BIOTECHNOLOGY (TIANJIN) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the bone marrow-derived mesenchymal stem cells have the following problems: with the increase of age, the number of stem cells and their ability to proliferate and differentiate significantly decrease; there is damage to the patient when the material is taken; it cannot be collected when there is a bone marrow disease; even if it is a healthy donor, it cannot extracting too much bone marrow
[0006] Although it has been confirmed that there are a large number of mesenchymal stem cells in the umbilical cord of newborns, the mesenchymal stem cells isolated from the umbilical cord will also undergo different degrees of differentiation during long-term culture in vitro. The operation of long-term culture and maintaining an undifferentiated state The technology is not mature

Method used

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  • Method for preparing mesenchymal stem cell
  • Method for preparing mesenchymal stem cell
  • Method for preparing mesenchymal stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1 (tissue block adherence method):

[0038] (1) Umbilical cord pretreatment

[0039] Take the fresh umbilical cord of the newborn, ligate both ends of the umbilical cord, put it into 100ml of RPMI-1640 culture solution containing 100U / ml penicillin and 100mg / ml streptomycin, and transport it at 4°C; Soak the umbilical cord in % medical alcohol for 45 seconds, remove about 2 cm from both ends of the ligated umbilical cord, cut the middle section into 1.5 cm long pieces, and obtain 20 pieces in total. Rinse the small section of umbilical cord repeatedly with normal saline to remove residual blood clots, cut open the small section of umbilical cord, remove the umbilical arterial vein, rinse repeatedly with normal saline until there is no residual blood, and cut into pieces to 1mm 3 , get a small piece of umbilical cord tissue with a volume of about 30ml, and refrigerate at 4°C;

[0040] (2) Cryopreservation of umbilical cord tissue blocks

[0041] Prepare 30m...

Embodiment 2

[0054] Embodiment 2 (enzyme digestion method):

[0055] (1) Umbilical cord pretreatment

[0056] Take the fresh umbilical cord of the newborn, ligate both ends of the umbilical cord, put it into 100ml of RPMI-1640 culture solution containing 100U / ml penicillin and 100mg / ml streptomycin, and transport it at 4°C; Soak the umbilical cord in % medical alcohol for 45 seconds, remove about 2 cm from both ends of the ligated umbilical cord, cut the middle section into 1.5 cm long pieces, and obtain 20 pieces in total. Rinse the small section of umbilical cord repeatedly with normal saline to remove residual blood clots, cut open the small section of umbilical cord, remove the umbilical arterial vein, rinse repeatedly with normal saline until there is no residual blood, and cut into pieces to 1mm 3 , get a small piece of umbilical cord tissue with a volume of about 30ml, and refrigerate at 4°C;

[0057] (2) Cryopreservation of umbilical cord tissue blocks

[0058] Take out 30ml of ...

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PUM

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Abstract

The invention belongs to the technical field of biology, and discloses a method for preparing mesenchymal stem cells. The method takes a neonatal umbilical cord as a raw material and comprises the following steps: pretreating the umbilical cord; cryopreserving tissues; resuscitating the tissues; separating primary mesenchymal stem cells; and amplifying the mesenchymal stem cells. The method disclosed by the invention has the advantages being easy for industrialization and operation and the like, and can be used for cryopreserving a large quantity of umbilical cord tissues and resuscitating the umbilical cord tissues for clinical need to prepare the mesenchymal stem cells. Compared with an existing method for directly separating the mesenchymal stem cells and cryopreserving the mesenchymal stem cells, the method disclosed by the invention can be used for sufficiently preserving the most original mesenchymal stem cells contained in the umbilical cord tissues, thereby greatly reducing the preservation cost and preventing the mesenchymal stem cells from influencing clinical application after being differentiated due to multiple-time passage in vitro.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing mesenchymal stem cells. Background technique [0002] With the rise of stem cell research, especially the in-depth study of adult stem cells, it has become possible to isolate and culture stem cells from different tissue sources in vitro and establish corresponding cell lines. Among them, breakthroughs have been made in researches on the in vivo distribution, in vitro isolation, culture, induced differentiation, and developmental potential of mesenchymal stem cells. Since mesenchymal stem cells are widely distributed in adult tissues, easy to obtain, and can be expanded and cultured in vitro under simulated conditions in vivo, the most important thing is that many of their clinical application potentials have been confirmed, such as in tissue engineering wounds, burns, etc. , ischemic necrosis, bone marrow injury and other repair processes, and has...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 陈义龙
Owner SHUNHAO CELL BIOTECHNOLOGY (TIANJIN) CO LTD
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