Serum-free complete culture medium for inducing mesenchymal stem cells to be differentiated into corneal epithelial cells

Abstract

The invention relates to the field of cell induction differentiation and in particular relates to a serum-free complete culture medium for inducing mesenchymal stem cells to be differentiated into corneal epithelial cells. The serum-free complete culture medium is prepared by the following steps: uniformly mixing 5-10[mu] mol of resveratrol, 2-4[mu] mol of icariin, 1-3nmol of aspirin, 1-3nmol of parathyroid hormone, 5-10nmol of hydrocortisone, 1-3mg of rapamycin, 2-10[mu] g of testosterone, 2-10[m] g of EPO (erythropoietin), 2-10[mu] g of LIF (laser induced fluorescence) and the balance of a corneal epithelial cell serum-free complete culture medium in every 1L of the induction differentiation serum-free complete culture medium, and performing filtering and sterilization, so as to obtain the serum-free complete culture medium. According to the serum-free complete culture medium for inducing mesenchymal stem cells to be differentiated into corneal epithelial cells, traditional Chinese medicine components, namely resveratrol and icariin and aspirin, parathyroid hormone, hydrocortisone, rapamycin, testosterone and growth factors are combined to synergistically induce the mesenchymal stem cells to be differentiated into the corneal epithelial cells, selected induction components are all non-toxic, high in induction efficiency and short in induction time, the activity of the cornealepithelial cells obtained through induction is good, no rejection is caused after cell transplanting, ethical issues can be avoided, and the security is high.

Description

technical field [0001] The invention relates to the field of induction and differentiation of stem cells, in particular to a serum-free complete medium for inducing the differentiation of mesenchymal stem cells into corneal epithelial cells. Background technique [0002] The corneal epithelial layer is located on the outer surface of the cornea. There are 5-6 layers of cells in the upper layer of the cornea, and each layer is arranged very neatly and tightly. The deepest layer is a single layer of short columnar basal cells; there are 2-3 layers of polygonal wing cells above the deep layer, and the top layer is 2-3 layers of polygonal superficial cells cell. The superficial layer of superficial cells is non-keratinized flat cells, the gap between epidermal cells is not clear, the superficial cell membrane is very smooth, the cytoplasm is clear and transparent, the cytoplasmic protrusions combine with each other to form bridges, and the nucleus still exists. The cells are e...

AI Technical Summary

Problems solved by technology

However, only about 10,000 cases of corneal transplantation can be carried out in my country every year. The main reason is that the number of donated corneas is seriously insufficient, which limits the popularization and application of the surgery, resulting in the vast majority of patients with corneal blindness unable to regain their sight through corneal transplantation. Reconstruction of tissue engineered cornea is an important breakthrough to solve the shortage of donor cornea materials

[0004] However, there are currently some bottlenecks in corneal tissue engineering, such as the source of corneal epithelial seed cells.

The study of corneal epithelium has enabled people to have a deeper understanding of the physiological and pathological characteristics of the cornea and corneal diseases. However, due to the short life cycle of the differentiated corneal epithelial cells in vitro, they can only be passed on for 2 to 3 generations. The quantity is small and the cost is high, which limits the research of corneal tissue and the construction of tissue engineered cornea

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