Universität zu Köln

Kappler, Benjamin, Anic, Petra, Becker, Matthias, Bader, Andreas, Klose, Kristin, Klein, Oliver, Oberwallner, Barbara, Choi, Yeong-Hoon, Falk, Volkmar ORCID: 0000-0002-7911-8620 and Stamm, Christof (2016). The cytoprotective capacity of processed human cardiac extracellular matrix. J. Mater. Sci.-Mater. Med., 27 (7). DORDRECHT: SPRINGER. ISSN 1573-4838

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Abstract

Freshly isolated human cardiac extracellular matrix sheets (cECM) have been shown to support stem cell proliferation and tissue-specific lineage commitment. We now developed a protocol for standardized production of durable, bio-functional hcECM microparticles and corresponding hydrogel, and tested its cytoprotective effects on contractile cells subjected to ischemia-like conditions. Human ventricular myocardium was decellularized by a 3-step protocol, including Tris/EDTA, SDS and serum incubation (cECM). Following snap-freezing and lyophilization, microparticles were created and characterized by laser diffraction, dynamic image analysis (DIA), and mass spectrometry. Moreover, cECM hydrogel was produced by pepsin digestion. Baseline cell-support characteristics were determined using murine HL-1 cardiomyocytes, and the cytoprotective effects of ECM products were tested under hypoxia and glucose/serum deprivation. In cECM, glycoproteins (thrombospondin 1, fibronectin, collagens and nidogen-1) and proteoglycans (dermatopontin, lumican and mimecan) were preserved, but residual intracellular and blood-borne proteins were also detected. The median particle feret diameter was 66 mu m (15-157 mu m) by laser diffraction, and 57 mu m (20-182 mu m) by DIA with crystal violet staining. HL-1 cells displayed enhanced metabolic activity (39 +/- 12 %, P < 0.05) and proliferation (16 +/- 3 %, P < 0.05) when grown on cECM microparticles in normoxia. During simulated ischemia, cECM microparticles exerted distinct cytoprotective effects (MTS conversion, 240 +/- 32 %; BrdU uptake, 45 +/- 14 %; LDH release, -72 +/- 7 %; P < 0.01, each). When cECM microparticles were solubilized to form a hydrogel, the cytoprotective effect was initially abolished. However, modifying the preparation process (pepsin digestion at pH 2 and 25 degrees C, 1 mg/ml final cECM concentration) restored the cytoprotective cECM activity. Extracellular matrix from human myocardium can be processed to yield standardized durable microparticles that exert specific cytoprotective effects on cardiomyocyte-like cells. The use of processed cECM may help to optimize future clinical-grade myocardial tissue engineering approaches.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Kappler, Benjamin UNSPECIFIED UNSPECIFIED UNSPECIFIED Anic, Petra UNSPECIFIED UNSPECIFIED UNSPECIFIED Becker, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Bader, Andreas UNSPECIFIED UNSPECIFIED UNSPECIFIED Klose, Kristin UNSPECIFIED UNSPECIFIED UNSPECIFIED Klein, Oliver UNSPECIFIED UNSPECIFIED UNSPECIFIED Oberwallner, Barbara UNSPECIFIED UNSPECIFIED UNSPECIFIED Choi, Yeong-Hoon UNSPECIFIED UNSPECIFIED UNSPECIFIED Falk, Volkmar UNSPECIFIED orcid.org/0000-0002-7911-8620 UNSPECIFIED Stamm, Christof UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-270946
DOI:	10.1007/s10856-016-5730-5
Journal or Publication Title:	J. Mater. Sci.-Mater. Med.
Volume:	27
Number:	7
Date:	2016
Publisher:	SPRINGER
Place of Publication:	DORDRECHT
ISSN:	1573-4838
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ISCHEMIA-REPERFUSION; SCAFFOLD; CARDIOMYOCYTES; PLATFORM; INJURY; MODEL; GEL Multiple languages Engineering, Biomedical; Materials Science, Biomaterials Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/27094