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Mesenchymal stem cell serum-free medium and cell isolation and cultivation methods

A technology of serum-free medium and mesenchymal stem cells, which is applied in the field of cell biology, can solve the problems of unfavorable clinical application of cells, complex medium components, and low separation success rate, and achieve the effects of short preparation time, low cost, and simple operation

Pending Publication Date: 2016-09-21
SICHUAN NEO LIFE STEM CELL BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing technology for obtaining mesenchymal stem cells from placenta and umbilical cord is still not perfect, and there are still disadvantages such as complex composition of the culture medium used for separation or containing animal-derived components such as fetal bovine serum, low separation success rate, and high separation cost. Isolated cells are not conducive to clinical application
For example, the patent application with the publication number CN103243071A discloses a medium for the isolation and cultivation of mesenchymal stem cells. Although it does not contain serum, the medium contains various vitamins, metal ions and other components, and the preparation is complicated, and high cost
The patent application whose publication number is CN103805562A also faces the same high cost problem

Method used

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  • Mesenchymal stem cell serum-free medium and cell isolation and cultivation methods
  • Mesenchymal stem cell serum-free medium and cell isolation and cultivation methods
  • Mesenchymal stem cell serum-free medium and cell isolation and cultivation methods

Examples

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Embodiment 1

[0044] The preparation of embodiment 1 serum-free medium of the present invention

[0045] Take 97mL of DMEM / F12 basal medium, add 2mL of human platelet lysate, 1mL of non-essential amino acids, filter aseptically, and store at 4°C.

Embodiment 2

[0046] The preparation of embodiment 2 serum-free medium of the present invention

[0047] Take 94mL of DMEM / F12 basal medium, add 5mL of human platelet lysate, 1mL of non-essential amino acids, filter aseptically, and store at 4°C.

Embodiment 3

[0048] Example 3 Isolation, culture and identification of mesenchymal stem cells of the present invention (placenta)

[0049] 1. Isolation and culture method

[0050] 1. Human placenta is collected (on the premise that the mother signs the informed consent, both cesarean section and normal delivery are acceptable, and no sterility is required), and it is transported in a low-temperature ice box.

[0051] 2. Placental chorionic plate sampling and inoculation:

[0052] 1) High-temperature disinfection of surgical instruments and large stainless steel dressing boxes, while ultraviolet disinfection of biological safety cabinets;

[0053] 2) Strict screening of donors, blood tests to exclude infectious diseases (including AIDS, hepatitis A, B, C and syphilis);

[0054]3) Soak the placenta in a large stainless steel dressing box filled with pre-cooled physiological saline. The following steps are all carried out under sterile conditions. After peeling off the amniotic membrane on ...

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Abstract

The invention provides a mesenchymal stem cell serum-free medium. The mesenchymal stem cell serum-free medium is prepared from, by volume, 94-97 parts of DMEM / F12, 2-5 parts of human blood platelet lysate and 1 part of non-essential amino acid. The invention further provides application of the medium and isolation and cultivation methods of the mesenchymal stem cell serum-free medium. The serum-free medium can be used for effectively separating and cultivating mesenchymal stem cells derived from placentae and umbilical cords, the effects are superior to commercially available media, the cost is low, the safety is high, and the application prospect is good.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a serum-free medium for mesenchymal stem cells and a method for separating and culturing cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a kind of pluripotent stem cells derived from mesoderm, which have high self-renewal and multilineage differentiation potential, and can differentiate into various types of tissue cells under specific induction conditions. Good seed cells for cell transplantation and tissue engineering, with very broad application prospects. [0003] According to the source, MSCs can be divided into two categories: one is MSCs derived from adult tissues, such as bone marrow and adipose tissue; the other is MSCs derived from fetal / perinatal tissues, such as placenta and umbilical cord. MSCs derived from adult tissues are difficult to mass-produce for clinical treatment due to their limited proliferation potential, while MSCs derived...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2500/90
Inventor 伍明俊陈瑶瑶周敏邹庆陈强
Owner SICHUAN NEO LIFE STEM CELL BIOTECH
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