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Cranial carotid artery dissection specific induced pluripotent stem cell line carrying FBN1 mutation

A technology of human pluripotent stem cells and pluripotent stem cells, which is applied in the field of biomedicine and can solve problems such as unretrieved

Pending Publication Date: 2021-11-30
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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Problems solved by technology

One of the patients’ peripheral blood cells was reprogrammed into patient-specific iPSCs using the episomal method. At present, there is no international patent application for a craniocarotid artery dissection-specific induced human pluripotent stem cell line carrying the c.1858C>T mutation of the FBN1 gene.

Method used

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  • Cranial carotid artery dissection specific induced pluripotent stem cell line carrying FBN1 mutation
  • Cranial carotid artery dissection specific induced pluripotent stem cell line carrying FBN1 mutation
  • Cranial carotid artery dissection specific induced pluripotent stem cell line carrying FBN1 mutation

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Embodiment

[0043] 1. A preparation method for craniocarotid artery dissection-specific induced pluripotent stem cells containing the c.1858C>T mutation of the FBN1 gene, comprising the following steps:

[0044] (1) PBMCs were extracted from the blood of a patient with craniocervical dissection in Huashan Hospital Affiliated to Fudan University as a sample. Put PBMCs into PBMC complete medium for recovery, inoculate into 24-well plates and culture for 3 days, half change fresh PBMC complete medium every day.

[0045] (2) Use CytoTune TM 2.0 Sendai virus kit was used to transduce the revived PBMC, after incubation overnight, the cells were collected and centrifuged to remove Sendai virus. Cells were resuspended in PBMC complete medium and transferred to 24-well plates for continuous culture for 2 days. On day 3, the medium was replaced with cytokine-free PBMC medium, and the transduced cells were plated on vitronectin-coated cell plates. Change the cytokine-free PBMC medium every other ...

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Abstract

The invention relates to a cranial carotid artery dissection specific induced pluripotent stem cell line carrying FBN1 gene c.1858C > T mutation, and belongs to the technical field of biological medicine. The cell line is the cranial carotid artery dissection specific human induced multifunctional stem cell line carrying FBN1 gene c.1858C > T mutation; the strain is preserved in the China Center for Type Culture Collection on September 1, 2021, and the preservation number of the strain is CCTCC NO: C2021229. The establishment of the cell line provides a cell model for family hereditary CAD caused by gene mutation; a cell model is provided for new drug screening and drug treatment mechanism research of clinical cranial carotid dissection diseases; cell resources of hereditary diseases are preserved, a disease-specific hiPSC cell bank is enriched, and storage is made for subsequent research.

Description

technical field [0001] The invention relates to a craniocarotid artery dissection-specific induced pluripotent stem cell line carrying FBN1 mutation, and belongs to the technical field of biomedicine. Background technique [0002] Craniocarotid artery dissection CAD (Cervicocephalic artery dissection, CAD) refers to the tearing of the intima and media or media and adventitia of intracranial and carotid arteries due to various reasons, resulting in blood entering the arterial wall through the tear Between the formation of intramural hematoma, if the formation of thrombus can lead to cerebral artery embolism or even occlusion. Craniocarotid dissection is one of the most common causes of ischemic stroke in young adults. Epidemiological studies have found that 10-25% of the first ischemic stroke patients under the age of 45 are caused by craniocervical artery dissection; in addition, because some arterial dissections do not cause any clinical symptoms or only cause mild non-spe...

Claims

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Application Information

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IPC IPC(8): C12N5/074C12Q1/6883C12Q1/02
CPCC12N5/0696C12Q1/6883G01N33/5005C12Q2600/156
Inventor 韩翔徐昳康小翠朱珠董强
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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