Efficient pre-treatment device for culturing human umbilical cord mesenchymal stem cells and related application thereof

Abstract

The invention discloses an efficient pre-treatment device for culturing human umbilical cord mesenchymal stem cells and a related application thereof. The efficient pre-treatment device comprises a turnover upper cover, a cutting assembly, a digestion assembly and a cleaning centrifugal part, wherein a cutter groove is formed in the upper cover; the cutting assembly located at an opening of a shell is composed of latticed blades and an elastic seal ring and the latticed blades and the cutter groove are correspondingly arranged; 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. 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 umbilical cord 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 belongs to the technical field of human cell culture, in particular to a high-efficiency pretreatment device for culturing human umbilical cord mesenchymal stem cells and its corresponding application Background technique

[0002] Mesenchymal stem cells are a kind of very characteristic adult stem cells. These cells have the potential to differentiate into a variety of mature cells including: adipocytes, chondrocytes, bone cells, tenocytes and muscle cells. These properties, as well as their developmental plasticity, have drawn considerable attention from the scientific community as they could be exploited in regenerative medicine research. In vitro, mesenchymal stem cells isolated from umbilical cord Wharton's jelly can differentiate into neurons and glial cells after being induced by nerve-directed induction medium, Shh and FGF8. Mesenchymal stem cells can also be differentiated into adipocytes and osteoblasts. In addition, in the my...

Examples

Embodiment 1

[0062] Example 1 as figure 1 As shown, this embodiment provides a high-efficiency pretreatment device for human umbilical cord mesenchymal stem cells, which specifically includes: a reversible upper cover 100, the upper cover 100 is provided with knife grooves 101, and the knife grooves 101 are composed of at least two groups Different depths and mutual grid-like sub-slots are composed; the housing 200 hingedly connected with the upper cover 100; the cutting assembly 300 located at the opening of the housing 200, the cutting assembly 300 is arranged in cooperation with the upper cover 100 to wash The final umbilical cord segment is cut into small pieces of tissue with the same size; the cutting assembly 300 is composed of a grid-shaped blade 301 and an elastic sealing ring 302, and the grid-shaped blade 301 is composed of at least two groups of sub-blades arranged in an upper and lower staggered arrangement, And the grid-shaped blade 301 is set corresponding to the knife groov...

Embodiment 2

[0076] Example 2 This example provides a method for pretreatment of human umbilical cord mesenchymal stem cells using the high-efficiency pretreatment device of Example 1, including the following steps:

[0077] Step 1) Acquisition of Human Umbilical Cord Tissue

[0078] To obtain human umbilical cord tissue from newborns, try to use newborn umbilical cords delivered by cesarean section to avoid contamination; put the umbilical cord segment in iced saline, keep it at a low temperature of about 4 degrees, and quickly bring it back to the cell laboratory. In the safety cabinet, the umbilical cord segment was fully washed with iced saline to remove blood and impurities. After the umbilical artery and umbilical vein were removed, the umbilical cord segment was repeatedly folded into 5-8 layers of 5 cm × 5 cm tissue blocks. Put the tissue block into the cutting assembly 300, select the cutting specifications of the grid-shaped blade 301 and the knife groove 101 according to the nee...

Embodiment 3

[0083] Example 3 This example provides a method for culturing human umbilical cord mesenchymal stem cells pretreated in Example 2 above, including the following steps:

[0084] Step 1) Culture medium preparation

[0085] 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; in order to improve human umbilical cord mesenchymal For the adherent survival rate of stem cells, on the first day of primary culture, subculture and recovery, culture with DMEM / F12 medium containing 20% ​​fetal bovine serum and 1% penicillin-streptomycin; at other times, culture with 10% fetal Bovine serum, 1% penicillin and streptomycin in DMEM / F12 medium for cultivation;

[0086] Step 2) Two-dimensional culture

[0087] The purer human umbilical cord mesenchymal stem cells pretreated in Example 2 were taken, and the purer human umbilical cord mesenchymal stem cells were resuspend...

technical field

[0001] The invention belongs to the technical field of human cell culture, in particular to a high-efficiency pretreatment device for culturing human umbilical cord mesenchymal stem cells and its corresponding application Background technique

[0002] Mesenchymal stem cells are a kind of very characteristic adult stem cells. These cells have the potential to differentiate into a variety of mature cells including: adipocytes, chondrocytes, bone cells, tenocytes and muscle cells. These properties, as well as their developmental plasticity, have drawn considerable attention from the scientific community as they could be exploited in regenerative medicine research. In vitro, mesenchymal stem cells isolated from umbilical cord Wharton's jelly can differentiate into neurons and glial cells after being induced by nerve-directed induction medium, Shh and FGF8. Mesenchymal stem cells can also be differentiated into adipocytes and osteoblasts. In addition, in the my...

Examples

Embodiment 1

[0062] Example 1 as figure 1 As shown, this embodiment provides a high-efficiency pretreatment device for human umbilical cord mesenchymal stem cells, which specifically includes: a reversible upper cover 100, the upper cover 100 is provided with knife grooves 101, and the knife grooves 101 are composed of at least two groups Different depths and mutual grid-like sub-slots are composed; the housing 200 hingedly connected with the upper cover 100; the cutting assembly 300 located at the opening of the housing 200, the cutting assembly 300 is arranged in cooperation with the upper cover 100 to wash The final umbilical cord segment is cut into small pieces of tissue with the same size; the cutting assembly 300 is composed of a grid-shaped blade 301 and an elastic sealing ring 302, and the grid-shaped blade 301 is composed of at least two groups of sub-blades arranged in an upper and lower staggered arrangement, And the grid-shaped blade 301 is set corresponding to the knife groov...

Embodiment 2

[0076] Example 2 This example provides a method for pretreatment of human umbilical cord mesenchymal stem cells using the high-efficiency pretreatment device of Example 1, including the following steps:

[0077] Step 1) Acquisition of Human Umbilical Cord Tissue

[0078] To obtain human umbilical cord tissue from newborns, try to use newborn umbilical cords delivered by cesarean section to avoid contamination; put the umbilical cord segment in iced saline, keep it at a low temperature of about 4 degrees, and quickly bring it back to the cell laboratory. In the safety cabinet, the umbilical cord segment was fully washed with iced saline to remove blood and impurities. After the umbilical artery and umbilical vein were removed, the umbilical cord segment was repeatedly folded into 5-8 layers of 5 cm × 5 cm tissue blocks. Put the tissue block into the cutting assembly 300, select the cutting specifications of the grid-shaped blade 301 and the knife groove 101 according to the nee...

Embodiment 3

[0083] Example 3 This example provides a method for culturing human umbilical cord mesenchymal stem cells pretreated in Example 2 above, including the following steps:

[0084] Step 1) Culture medium preparation

[0085] 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; in order to improve human umbilical cord mesenchymal For the adherent survival rate of stem cells, on the first day of primary culture, subculture and recovery, culture with DMEM / F12 medium containing 20% ​​fetal bovine serum and 1% penicillin-streptomycin; at other times, culture with 10% fetal Bovine serum, 1% penicillin and streptomycin in DMEM / F12 medium for cultivation;

[0086] Step 2) Two-dimensional culture

[0087] The purer human umbilical cord mesenchymal stem cells pretreated in Example 2 were taken, and the purer human umbilical cord mesenchymal stem cells were resuspend...

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