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

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

CN105950550APending Publication Date: 2016-09-21SICHUAN NEO LIFE STEM CELL BIOTECH
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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

Experimental program
Comparison scheme
Effect test

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

Patent Timeline
21 Sep 2016
Publication
CN105950550A
IPC
C12N5/0775
CPC
C12N5/0668; C12N2500/90
Inventors
伍明俊; 陈瑶瑶