Cell and drug targeting transfer structure as well as preparation method and application thereof
A cell and transfer technology, applied in the field of cell therapy and immunotherapy of diseases, can solve the problems of complex operation and high cost
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Embodiment 1
[0130] Liposomes are artificial bilayer vesicles formed by lipid molecules. The structure of a lipid molecule usually consists of three parts: a hydrophilic head, a hydrophobic tail and a connecting part. In the water medium, due to the hydrophobic interaction, the hydrophobic tails of the lipid molecules are close to each other, and the hydrophilic heads are oriented toward the water phase to form a bilayer. Based on such a structure, hydrophilic drugs can be entrapped in the inner water phase of liposome (inner aqueous phase), and hydrophobic drugs can also be embedded in the lipid bilayer membrane.
[0131] 1. Preparation of HSPC / CHOL ordinary lipid nanoparticles and fluorescent HSPC / CHOL lipid nanoparticles
[0132] 1) Dissolve 5.9mg of hydrogenated soybean lecithin (HSPC), 2.1mg of cholesterol (Chol), and 1.2mg of DSPE-PEG (2000) in 1ml of chloroform. If fluorescent lipid nanoparticles are prepared, additional DiI fluorescent dye chloroform is required for storage Solut...
Embodiment 2
[0183] Example 2 gamma delta T cells are effector cells
[0184] Preparation of Zoledronic acid liposome preparations by lyophilization and hydration:
[0185] Weigh the EPC / HSPC / PA and Chol in the ratio of the prescription amount, dissolve it in tert-butanol and put it in a freeze-drying bottle, heat it in a water bath at 70 degrees to fully mix and dissolve, and then freeze-dry it in a freeze dryer for 24 hours to obtain the ratio of each recipe The lyophilized powder is added to zoledronic acid for hydration when used, and the pH is adjusted to about 7-8, and it is passed through a 400nm, 200nm, 100nm, 80nm polydin film extrusion device in turn under heating in a 70-degree water bath to control the particle size. Liposomes with a diameter of about 120 nm were obtained, and the free drug solution that was not encapsulated in the liposomes was separated by ultracentrifugation.
[0186]Take an appropriate amount of anti-cluster of differentiation 3 single-chain antibody (anti...
Embodiment 3
[0192] Example 3 Induced pluripotent stem cells are effector cells
[0193] Induced pluripotent stem cells (IPS cells) are increasingly used in biological therapy. They are pluripotent stem cells formed by the dedifferentiation of mammalian adult cells through transduction of transcription factors. We added Nano liposomes are connected with LGR5 antibody fragments and transferrin receptor antibody fragments respectively, and the GSK-2 inhibitor CHIR99021 is loaded in the liposomes.
[0194] Preparation of CHIR99021 liposome preparation by ethanol injection method: EPC / Chol / CHIR99021 was dissolved in ethanol at a ratio of 6:2:1, heated in a 70-degree water bath to fully mix and dissolve, slowly dripped into the aqueous solution, and adjusted to a pH of about 7-8. Under heating in a 70-degree water bath, the liposomes are passed through 200nm, 100nm, and 50nm membrane extruders sequentially to control the particle size to obtain liposomes of about 100nm.
[0195] Take an approp...
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