Mesenchymal stem cell particles

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

Inactive Publication Date: 2014-08-20
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] Protocols for MSC generation and CM preparation have been described previously 15，16 .

[0375]Briefly, chemically defined serum-free media were conditioned by MSCs derived from human embryonic stem cells (hESCs) using a clinically compliant protocol. Three polyclonal, karyotypically stable, and phenotypically MSC-like cultures were generated by trypsinization and propagation of hESCs from either the HuES9 hESC line or the H1 hESC line in feeder-free and serum-free selection media, which did not Expresses pluripotency-associated markers but exhibits 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 for at least 80 population doublings. To harvest MSC secretions, hESC-derived MSC cultures were transferred to chemically defined serum-free medium to condition the medium for 3 day...

Embodiment 2

[0378] Example 2: Materials and Methods: Animals

[0379] All experiments were performed in accordance with the "Guide for the Care and Use of Laboratory Pigs" prepared by the Institute of Laboratory Animal Resources and in accordance with the Animal Experimentation Committee of the Faculty of Pharmacy, Utrecht University, The Netherlands. implementation of prior approval.

Embodiment 3

[0380] Example 3: Materials and Methods: Study Design

[0381] 30 female Dalland Landrace pigs (60-70 kg; IDDLO, Lelystad, Netherlands) (both pretreated with clopidogrel 75 mg / day for 3 days and amiodarone 400 mg / day for 10 days) were randomized Classified as MSC-CM, non-CM, or saline treatment.

[0382] A saline group was added to assess the potential effect of fresh, non-conditioned 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 min ischemic period was chosen to cause severe myocardial damage without inducing completely transmural myocardial infarction. Use a 4-h reperfusion period because infarct size measurements using TTC staining are most reliable after 3 h of reperfusion 17 . After longer periods of reperfusion, it becomes more difficult to assess the state of oxidative stress and mechanisms of apoptosis.

[0383] Treatment was initiated 5 minutes befor...

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Abstract

The present invention describes a particle secreted by mesenchymal stem cells and comprising at least one biological characteristic of mesenchymal stem cells. The biological properties may comprise biological activities of mesenchymal stem cell conditioned medium (MSC-CM), such as cardioprotection or reduction in infarct size. The particles may comprise vesicles or exosomes.

Description

field of invention [0001] The present invention relates to the fields of development, cell biology, molecular biology and genetics. More specifically, the present invention relates to a method of deriving mesenchymal stem cells from embryonic stem cells. Background of the invention [0002] Unlike already differentiated cells, stem cells have the ability to divide and self-renew or differentiate into phenotypically and functionally distinct 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 (MSCs) are pluripotent stem cells with documented evidence of therapeutic efficacy in treating musculoskeletal injuries, improving cardiac function in cardiovascular disease and ameliorating the severity of GVHD (Le Blanc and Pittenger, 2005). Lineage-restricted, they have a limited but strong potential ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/12C12N5/06A61K35/28C12N5/0775
CPCA61K35/12A61K35/28C12N5/0663A61P3/10A61P9/00A61P9/04A61P9/10A61P11/02A61P13/12A61P17/00A61P17/02A61P17/06A61P17/12A61P19/08A61P25/16A61P25/28A61P29/00A61P35/00A61P37/02A61P37/04A61P37/08C12N5/00C12N5/0667A61K9/0019A61K9/10A61K47/24A61K47/28A61L2300/30
Inventor 林赛娟
Owner AGENCY FOR SCI TECH & RES
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