Method for construction of stem cell in-vitro multiplication and differentiation microenvironment

Abstract

The invention discloses a constructing method of stem cell breeding and differentiation microenvironment in vitro in the stem cell tissue engineering domain, which comprises the following steps: adopting sodium alginate and polylysine as material to prepare APA microencapsulation ES cell; adding microencapsulation ES cell in the ES growing culture medium; stewing at 37 Deg C in the culture box with 5% CO2; changing culture medium every 2-3d to obtain the product. The invention simplifies the operation to generate high-density and large scale augmentation, which can inhibit or delay the oriental differentiation for ES cell.

Description

technical field [0001] The invention relates to the field of stem cell tissue engineering, in particular to a method for constructing a microenvironment for proliferation and differentiation of stem cells in vitro. Background technique [0002] Embryonic stem cells (ESC) is a specific cell group derived from the inner cell mass of the blastocyst, which has the potential of self-renewal and omnidirectional differentiation, that is, under suitable in vivo or in vitro conditions, it can differentiate into Cells of three different germ layers: inner, middle, and outer. In 1981, the successful isolation and establishment of mouse ES cells opened the prelude to stem cell research [Document 1: Evans MJ, Kaufman MH. Establishment in culture of pluripotential cells from mouseembryos. Nature, 1981; 292(5819): 154-156 .]. Subsequently, the directional induction of ESC differentiation has become a hot spot of common concern at home and abroad. At present, the research on the directio...

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Problems solved by technology

However, the above-mentioned approaches have the following problems: (1) The commodity price of cytokine LIF is expensive, which not only increases the harshness of ES cell maintenance culture conditions, but also greatly limits the extensive development of large-scale expansion of ES cells

(2) The preparation of feeder cells has high requirements on the source of materials (pregnant mice at 13.5 days pregnant), the operation is complicated, and feeder cells pretreated with mitomycin are required for each passage

(3) The transgenic operation of eukaryotic cells is relatively complicated, and the expression of foreign genes is unstable and easy to be lost in the process of proliferation and passage

However, in recent years, most of the relevant reports have focused on the in vivo induced differentiation of adult stem cells, and there are few studies on ES cells.

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