TY  - JOUR
A1  - Hackethal, Johannes
A1  - Dungel, Peter
A1  - Teuschl, Andreas Herbert
T1  - Frequently Used Strategies to Isolate Extracellular Matrix Proteins from Human Placenta and Adipose Tissue
JF  - Tissue Engineering Part C: Methods
N2  - The natural extracellular matrix (ECM) provides the optimal environment for cells. Many enzymatic or non-enzymatic based strategies to extract ECM proteins from tissues were published over the past years. However, every single isolation strategy reported so far is associated with specific bottlenecks. In this study, frequently used strategies to isolate ECM from human placenta or adipose tissue using Tris-, serum-, or pepsin-based buffers were compared. The resulting ECM proteins were biochemically characterized by analysis of cellular remnants using Hoechst DNA staining, glycosaminoglycan (GAG) content by dimethylmethylene blue, visualization of protein bands using sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis combined with amino acid quantification, and assessment of the proangiogenic profile using an angiogenesis array. Tris-NaCl-extracted ECM proteins showed a high heterogenic degree of extracted proteins, bioactive growth factors, and GAGs, but no collagen-I. Active serum-extracted ECM showed significant lower DNA remnants when compared with the Tris-NaCl isolation strategy. Pepsin-extracted ECM was rich in collagen-I and low amounts of remaining bioactive growth factors. This strategy was most effective to reduce DNA amounts when compared with the other isolation strategies. Pepsin-extracted ECM from both tissues easily gelled at 37°C, whereas the other extracted ECM strategies did not gel at 37°C (Tris-NaCl: liquid; serum: sponge). All relevant characteristics (DNA residues, ECM diversity and bioactivity, shape) of the extracted ECM proteins highly depend on its isolation strategy and could still be optimized. Impact statement The natural human extracellular matrix (ECM) is the ideal cell niche. Various strategies were reported to isolate human ECM components from various sources. In this article, we compared frequently used methods and compared their characteristics (DNA remnants, glycosaminoglycan content, sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, amino acid quantification, angiogenesis array, and gel formation). We conclude that more research is still necessary to optimize current isolation approaches for in vitro or in vivo applications of human ECM.
KW  - Tissue Engineering
KW  - Biomaterials
KW  - Adipose Tissue
KW  - extracellular matrix
KW  - human placenta
Y1  - 
VL  - 27
IS  - 12
SP  - 649
EP  - 660
ER  - 
TY  - CHAP
A1  - Hackethal, Johannes
A1  - Schuh, Christina
A1  - Hofer, Alexandra
A1  - Meixner, Barbara
A1  - Hennerbichler, Simone
A1  - Redl, Heinz
A1  - Teuschl, Andreas
T1  - Human Placenta Laminin-111 as a Multifunctional Protein for Tissue Engineering and Regenerative Medicine
T2  - Advances in Experimental Medicine and Biology
KW  - Biomaterial
KW  - Tissue Engineering
KW  - Regenerative medicine
Y1  - 
PB  - Springer
ER  - 
TY  - JOUR
A1  - Hackethal, Johannes
A1  - Mühleder, Severin
A1  - Hofer, Alexandra
A1  - Schneider, Karl Heinrich
A1  - Prüller, Johanna
A1  - Hennerbichler, Simone
A1  - Redl, Heinz
A1  - Teuschl, Andreas
T1  - An Effective Method of Atelocollagen Type 1/3 Isolation from Human Placenta and Its In Vitro Characterization in Two-Dimensional and Three-Dimensional Cell Culture Applications
JF  - Tissue Eng Part C Methods
KW  - Placenta
KW  - In Vitro
KW  - Cell Culture
Y1  - 2018
VL  - 23
IS  - 5
SP  - 274
EP  - 285
ER  -