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Mesenchymal stem cell particles

A technology of stem cells and particles, used in the fields of development, cell biology, molecular biology and genetics, to solve problems such as unmet needs

Inactive Publication Date: 2011-04-13
AGENCY FOR SCI TECH & RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Unmet need for 'existing' MSC-based treatment options (at affordable cost, with better quality control and consistency)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0373] Example 1: Materials and methods: MSC-CM preparation

[0374] The protocol for MSC generation and CM preparation has been described previously 15，16 .

[0375] In short, a clinical compliance protocol was used to condition a serum-free medium determined by the chemical composition of MSCs derived from human embryonic stem cells (hESC). Three types of polyclonal, stable karyotype and phenotypic MSC-like cultures were generated by trypsinization and propagation of hESC from HuES9 hESC line or H1 hESC line in feeder-free and serum-free selective medium. Express pluripotency-related markers, but display MSC-like surface antigens (CD29+, CD44+, CD49a+ / e+, CD105+, CD166+, CD34-, CD45-) and gene expression profiles 15 .

[0376] One of these cultures, HuES9.E1, can be stably expanded by at least 80 population doublings. In order to harvest MSC secretions, the hESC-derived MSC culture was transferred to a chemically defined serum-free medium to conditioned the medium for 3 days, aft...

Embodiment 2

[0378] Example 2: Materials and methods: animals

[0379] All experiments are in accordance with the "Guide for the Care and Use of Laboratory Pigs" prepared by the Institute of Laboratory Animal Resources (Institute of Laboratory Animal Resources) and in accordance with the Animal Experiment Committee of the School of Pharmacy, Utrecht University, the Netherlands The prior approval of implementation.

Embodiment 3

[0380] Example 3: Materials and methods: research design

[0381] Thirty female DallandLandrace pigs (60-70kg; IDDLO, Lelystad, Netherlands) (all pretreated with clopidogrel 75mg / day for 3 days and amiodarone 400mg / day for 10 days) were randomly selected Classified as MSC-CM, non-CM, or saline treatment.

[0382] The saline group was added to evaluate the potential effect of fresh, unconditioned media. In all pigs, MI was induced by ligation of the proximal left circumflex coronary artery (LCxCA) for 75 minutes followed by 4 hours of reperfusion. The 75-minute period of ischemia was chosen to cause severe myocardial damage without inducing completely transmural myocardial infarction (completely transmural myocardial infarction). Use a 4-hour reperfusion period because the infarct size measurement using TTC staining is the most reliable after 3 hours of reperfusion 17 . After a longer period of reperfusion, it becomes more difficult to assess the oxidative stress state and the me...

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Abstract

We describe a particle secreted by a mesenchymal stem cell and comprising at least one biological property of a mesenchymal stem cell. The biological property may comprise a biological activity of a mesenchymal stem cell conditioned medium (MSC- CM) such as cardioprotection or reduction of infarct size. The particle may comprise a vesicle or an exosome.

Description

Invention field [0001] The invention relates to the fields of development, cell biology, molecular biology and genetics. More specifically, the present invention relates to a method for deriving mesenchymal stem cells from embryonic stem cells. Background of the invention [0002] Different from differentiated cells, stem cells have the ability to divide and self-renew or differentiate into phenotypically and functionally different daughter cells (Keller, Genes Dev. 2005; 19:1129-1155; Wobus and Boheler, Physiol Rev. 2005 ; 85: 635-678; Wiles, Methods in Enzymology. 1993; 225: 900-918; Choi et al., Methods Mol Med. 2005; 105: 359-368). [0003] Mesenchymal stem cells (MSC) are pluripotent stem cells, which have written evidence of therapeutic efficacy in treating musculoskeletal injuries, improving heart function in cardiovascular diseases, and improving the severity of GVHD (Le Blanc and Pittenger, 2005). Due to lineage limitations, they have a limited but strong potential to di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/12C12N5/06A61K35/28C12N5/0775
CPCC12N5/0663A61K35/28A61K35/12A61P3/10A61P9/00A61P9/04A61P9/10A61P11/02A61P13/12A61P17/00A61P17/02A61P17/06A61P17/12A61P19/08A61P25/16A61P25/28A61P29/00A61P35/00A61P37/02A61P37/04A61P37/08C12N5/00C12N5/0667A61K9/0019A61K9/10A61K47/24A61K47/28A61L2300/30
Inventor 林赛娟
Owner AGENCY FOR SCI TECH & RES
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