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In vitro construction method for salivary glands organs and acinus

A construction method and salivary gland technology, which are applied in the field of in vitro construction of salivary gland organoids and acinar-like vesicles to achieve the effects of extensive sources, minimal trauma, and easy excision

Inactive Publication Date: 2015-09-02
赵振民
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Problems solved by technology

[0008] There is no study to obtain human salivary gland organoids by mixed three-dimensional culture of human minor salivary gland adult mesenchymal stem cells and human minor salivary gland adult epithelial stem / progenitor cells, or simply use human minor salivary gland adult epithelial stem / progenitor cells for three-dimensional culture to obtain human salivary gland organoids. A Study Report on Human Salivary Gland Acinar Tissue

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  • In vitro construction method for salivary glands organs and acinus
  • In vitro construction method for salivary glands organs and acinus
  • In vitro construction method for salivary glands organs and acinus

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Embodiment 1

[0022] Methods and reagents used in the culture process:

[0023] (1) Culture of primary cells: DMEM / F12 supplemented with 10% fetal bovine serum, 1% double antibody and 1% glutamine;

[0024] (2) In vitro expansion:

[0025] 1) Human minor salivary gland mesenchymal stem cell culture: Mesenchymal stem cell medium: DMEM / F12 supplemented with 5% fetal bovine serum, 1% double antibody, and BSA 10μg / mL, apo-transferrin 10μg / mL, insulin 5mg / mL mL, EGF 2ng / mL, FGF-2 2ng / mL and hydrocortisone 1mg / mL.

[0026] 2) Human minor salivary gland epithelial stem / progenitor cell culture: keratinocyte medium

[0027] DMEM / F12 is supplemented with BSA (bovine serum albumin) 5μg / mL, transferrin (transferrin) 5μg / mL, BPE (bovine pituitary extract) 50μg / mL, insulin (insulin) 3.75g / mL, FGF-2 3ng / mL, EGF 1ng / mL, epinephrine (epinephrine) 500ng / mL, hydrocortisone (hydrocortisone) 0.5g / mL, Prostaglandin E2 (prostaglandin E2) 10 -8 M and T3 30 nM.

[0028] 3) Salivary gland organoid construction...

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Abstract

The invention discloses an in vitro construction method for biological engineering salivary glands organs and acinus based on human minor salivary gland adult stem cells belonging to the technical field of tissue and organ reconstruction. The in vitro construction method specifically is a method for obtaining organoids with salivary gland structures and functions by mixed cultivation of mesenchymal stem cells and epithelial stem / progenitor cells of the human minor salivary glands, or a method for obtaining acinar-like tissues with salivary gland structures and functions by individual cultivation of epithelial stem / progenitor cells of the human minor salivary glands. The method has the advantages of being wide in sampling source, easy to cut, and small in trauma; a donor site in a mouth is private; materials are drawn from adult tissues; and autoplastic transplantation can be carried out through amplification in vitro. The method has a wide application prospect in growth and development research of in vitro or in vivo salivary glands, research on a salivary gland disease model, research on physiological functions and pathological changes of the saliva and treatment of salivary gland diseases.

Description

technical field [0001] The invention belongs to the technical field of tissue and organ reconstruction, and in particular relates to an in vitro construction method for bioengineered salivary gland organoids and acini-like glands based on human minor salivary gland adult stem cells. Background technique [0002] Studies have confirmed that major salivary gland tissue contains mesenchymal stem cells and epithelial stem cells. There are three major salivary glands in human salivary glands and many small salivary glands distributed in the oral mucosa. They are all composed of parenchyma (acini, ducts) and interstitium (connective tissue, blood vessels, lymphatic vessels and nerves, etc.). Research on stem cells in salivary glands mainly focuses on major salivary glands. It has been confirmed that mesenchymal stem cells exist in human parotid glands and can be induced to differentiate into osteogenic, adipogenic and chondrogenic cells in vitro. This becomes a new source of mese...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/074C12N5/071A61L27/38
CPCA61L27/38
Inventor 赵振民吕璘张辰刘磊韩婷璐张翔宇杜水果
Owner 赵振民
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