The isolated colonic mucosa: a new substrate for tissue reconstruction. Isolation, decellularization and characterization by immunohistochemistry and electron microscopy
Eur. j. anat
; 22(1): 1-15, ene. 2018. ilus, graf
Artigo
em Inglês
| IBECS
| ID: ibc-170476
Biblioteca responsável:
ES1.1
Localização: BNCS
ABSTRACT
This study proposes the use of isolated colonic mucosa as a "scaffold" for cell cultures and potentially for tissue reconstruction. The main goal of this study was to obtain complete decellularization of the specimens while preserving the superficial basement membrane (BM) as a place for cell attachment and growth. This decellularization technique uses a chelating agent in combination with mechanical vibration, followed by detergents and DNase. The grade of decellularization achieved is assessed by counting the number of cell nuclei stained with propidium iodide (PI). BM marker proteins such as collagen IV, laminin and perlecan were detected by immunohistochemistry (ICC). Transmission (TEM) and scanning electron microscopy (SEM) were used to examine the BM ultrastructure and surface topography. The results show that the protocol used is suitable for rat colonic mucosa. During the process, material of the lamina lucida (LL) was partly removed from the BM, whereas the lamina densa (LD) seems to have remained unchanged. The BM had become thinner than the control specimens. The nanotopography of the BM surface is characterized by globules of 34-60 nm in diameter. Human fetal fibroblasts were successfully cultured on this substrate confirming that cells can adhere to and grow on this substrate, at least for the particular cell line used. It can be said that the colonic mucosa is an interesting substrate for in vitro stud-ies with cells and presumably also for tissue reconstruction
RESUMEN
No disponible
Texto completo:
Disponível
Coleções:
Bases de dados nacionais
/
Espanha
Base de dados:
IBECS
Assunto principal:
Membrana Basal
/
Imuno-Histoquímica
/
Mucosa Intestinal
/
Mucosa
Limite:
Animais
Idioma:
Inglês
Revista:
Eur. j. anat
Ano de publicação:
2018
Tipo de documento:
Artigo
Instituição/País de afiliação:
Saarland University/Germany
/
University Rey Juan Carlos/Spain