Method for constructing cell transplantation slice by using human umbilical cord mesenchymal stem cells (hUC-MSCs) and amniotic membrane

A technique for stem cell and cell transplantation, which is applied in the field of human umbilical cord mesenchymal stem cells and amniotic membrane-constructed cell transplantation sheets, can solve problems such as poor effect, and achieve the effects of convenient clinical use, increased feasibility, and increased operability.

Inactive Publication Date: 2016-06-08
秦方园 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the main ways of clinical application of stem cells include intravenous reinfusion, injection of lesion site, lumbar puncture injection, catheter intervention, etc. These Approaches Are Less Effective Where Location Matters

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A method of using hUC-MSCs and amnion to construct a cell transplant sheet, comprising the steps of:

[0035] (1) Take freshly collected human amniotic membrane and umbilical cord, wash and soak the umbilical cord in normal saline for later use;

[0036] (2) Amniotic membrane treatment: bluntly separate the chorion from the cleaned amniotic membrane, then wipe the amniotic membrane with gauze moistened with normal saline, and then soak the amniotic membrane in normal saline containing penicillin 150U / ml and streptomycin sulfate 150U / ml After soaking for 0.5 hours, put it into sterile glycerin and dehydrate at 4°C. The dehydration time does not exceed 12 hours, and then transfer it to sterile glycerin and store it at -20°C for later use;

[0037] (3) Cut the soaked umbilical cord into small pieces with a length of about 1-2 cm, wash, then transfer the umbilical cord tissue to a petri dish, add physiological saline to the petri dish until 1 / 2 of the umbilical cord tissue ...

Embodiment 2

[0053] A method of using hUC-MSCs and amnion to construct a cell transplant sheet, comprising the steps of:

[0054] (1) Take freshly collected human amniotic membrane and umbilical cord, wash and soak the umbilical cord in normal saline for later use;

[0055] (2) Amniotic membrane treatment: bluntly separate the chorion from the cleaned amniotic membrane, then wipe the amniotic membrane with a gauze moistened with normal saline, and then soak the amniotic membrane in normal saline containing 200 U / ml penicillin and 200 U / ml streptomycin sulfate After immersing in sterile glycerin for 1 hour, put it into sterile glycerin and dehydrate at 8°C. The dehydration time should not exceed 12 hours, then transfer it to sterile glycerin and store it at -20°C for later use;

[0056] (3) Cut the soaked umbilical cord into small pieces with a length of about 1-2 cm, wash, then transfer the umbilical cord tissue to a petri dish, add physiological saline to the petri dish until 1 / 2 of the u...

Embodiment 3

[0065] A method of using hUC-MSCs and amnion to construct a cell transplant sheet, comprising the steps of:

[0066] (1) Take freshly collected human amniotic membrane and umbilical cord, wash and soak the umbilical cord in normal saline for later use;

[0067] (2) Treatment of amniotic membrane: bluntly separate the chorion from the cleaned amniotic membrane, then wipe the amniotic membrane with gauze moistened with normal saline, and then soak the amniotic membrane in normal saline containing 50 U / ml penicillin and 50 U / ml streptomycin sulfate After soaking for 0.5-1 hour, put it into sterile glycerin and dehydrate at 2°C, the dehydration time should not exceed 12 hours, then transfer it to a sterile glycerin bottle and store it at -20°C for later use;

[0068](3) Cut the soaked umbilical cord into small pieces with a length of about 1-2 cm, wash, then transfer the umbilical cord tissue to a petri dish, add physiological saline to the petri dish until 1 / 2 of the umbilical co...

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Abstract

The invention relates to a method for constructing a cell transplantation slice by using human umbilical cord mesenchymal stem cells (hUC-MSCs) and anthropogenic amniotic membrane. The method comprises the following steps: taking the freshly collected anthropogenic amniotic membrane and umbilical cord, separating chorion from the amniotic membrane, treating the separated amniotic membrane by normal saline, and then cryopreserving for later use; cutting the umbilical cord into small pieces, and tearing Wharton's jelly off; cutting the Wharton's jelly into pieces, putting the cut Wharton's jelly onto a sterile culture vessel, adding a culture solution into the sterile culture vessel, and putting the sterile culture vessel into an incubator for culturing to obtain primary generation hUC-MSCs; adding pancreatin into a culture medium of the primary generation hUC-MSCs, digesting, and continuously culturing in the incubator to obtain the hUC-MSCs; carrying out sterile hydration on the cryopreserved standby amniotic membrane, then flatly laying the amniotic membrane on an amniotic membrane carrier thimble, and then moving into the sterile culture vessel; inoculating the hUC-MSCs in the culture vessel, and then culturing in the incubator to obtain the cell transplantation slice with cell confluence reaching up to 80-90%. The cell transplantation slice prepared by the method is capable of carrying out damage repair at specific parts, so that the defects of using the hUC-MSCs or amniotic membrane alone for treatment are overcome; therefore, a new treatment strategy is provided for refractory diseases, and a new therapy approach is opened up for the clinical application of stem cells.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for constructing a cell transplant sheet from human umbilical cord mesenchymal stem cells and amniotic membrane. Background technique [0002] Stem cells are a class of cells with different differentiation potentials that self-renew in an undifferentiated state. Stem cell therapy refers to the process of using human autologous or allogeneic stem cells to infuse (or implant) into the human body after extracorporeal manipulation for disease treatment. Stem cells used in cell therapy mainly include adult stem cells, embryonic stem cells and induced pluripotent stem cells. At present, a number of clinical application studies of stem cells have been carried out at home and abroad, involving various types of stem cells and various types of diseases. The main disease types include bone and joint diseases, liver cirrhosis, graft host rejection (GVHD), spinal cord injury ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/073
CPCC12N5/0697C12N2502/025C12N2502/1388
Inventor 秦方园翟亚萍王丽娅
Owner 秦方园
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