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A stem cell culture system and a culture method thereof

A stem cell culture and stem cell technology, applied in the field of stem cell culture, can solve the problem of neural stem cell loss of traits, etc., and achieve the effects of promoting proliferation, improving biological safety, and facilitating separation

Inactive Publication Date: 2016-12-21
浙江译美生物科技有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

[0004] However, this method of culturing neural stem cells with high cell viability has the following disadvantages: suspension and adherent culture are alternately carried out in the culture medium, and the environment of neural stem cells is far from the state of natural growth in vivo, resulting in in vitro culture. The neural stem cells gradually lost their original traits in the culture room, and their shape, structure and function were far from their natural growth state in the body.

Method used

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  • A stem cell culture system and a culture method thereof

Examples

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Embodiment 1

[0034] Embodiment 1: A culture method of a stem cell culture system, comprising a), preparing a layer of feeder cells in a culture container, commercially purchased MEF cells, configuring MEF growth medium, and preparing it with high-glucose medium and 10% FCS; Prepare a 15ml centrifuge tube, add 5ml preheated MEF growth medium, take a MEF cell frozen in liquid nitrogen, shake gently in a water bath at 37 degrees Celsius, transfer the cells to the centrifuge tube after melting; collect the cells after centrifugation, and suspend Inoculate a 150mm tissue culture dish in 10ml of MEF growth medium, add MEF growth medium to 25ml, and incubate at 37°C until the cells are confluent. After the cells were confluent, mitomycin C was added, the medium was sucked off, and a freshly prepared MEF growth medium containing 10 ug / ml mitomycin C was added. Incubate at 37°C for 2.5 hours, in which mitomycin C stock solution, 2ml mitomycin C is dissolved in 5ml sterile PBS. The mother solution ...

Embodiment 2

[0040] Embodiment 2: A culture method of a stem cell culture system. The difference from Example 1 is that the components of the complete medium are configured according to the components in Table 1 below, and the results are displayed in the Listed in Table 1.

Embodiment 3

[0041] Embodiment 3: A culture method of a stem cell culture system. The difference from Example 1 is that the components of the complete medium are configured according to the components in the following Table 1, and the results are displayed in the Listed in Table 1.

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Abstract

A stem cell culture system is disclosed. The system comprises culture-layer cells, a cell sieve and a three-dimension medium in order from bottom to top. The three-dimension medium comprises a three-dimension support prepared from Matrigel and a complete medium. The complete medium comprises a DMEM medium, FCS, beta-mercaptoethanol, LIF, glutamine and insulin. The system is advantageous in that the system adopts the Matrigel and growth factors secreted from culture cells to stimulate an extracellular matrix wherein the stem cells in vivo are in, thus allowing an in-vitro living environment to be similar to an in-vivo living environment and ensuring proliferation and a normal form of the cells; stem cell proliferation is promoted through combined action of the culture-layer cells, the beta-mercaptoethanol and the insulin; stem cell differentiation during proliferation is inhibited through synergistic effects of LIF; and effects of capability of stimulating the in-vivo cytoplasmic matrix, high stem cell activity, safety and convenient separation are achieved.

Description

technical field [0001] The invention relates to the field of stem cell culture, in particular to a stem cell culture system and a culture method thereof. Background technique [0002] Stem cells are a type of pluripotent cells with self-replicating ability. Under certain conditions, it can differentiate into a variety of functional cells. Stem cells are divided into embryonic stem cells and adult stem cells according to their developmental stage. According to the developmental potential of stem cells, they are divided into three categories: totipotent stem cells, pluripotent stem cells and unipotent stem cells. Stem cells are not fully differentiated and immature cells, which have the potential to regenerate various tissues and organs and the human body, and are called "universal cells" in the medical field. [0003] The Chinese patent "A Method for Cultivating Neural Stem Cells Maintaining High Cell Viability" with the publication number CN103305466A discloses a method f...

Claims

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Application Information

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IPC IPC(8): C12N5/074C12N5/0735
CPCC12N5/0606C12N2500/38C12N2500/44C12N2501/33C12N2501/91C12N2513/00
Inventor 汪铮吴振化
Owner 浙江译美生物科技有限公司
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