Basics of isolation and cultivation of chondrocytes according to good laboratory practice
Metadatos
Mostrar el registro completo del ítemAutor
Muñoz Olmedo, Jose Manuel; Harvanová, Denisa; Špaková, Timea; Rosocha, Ján; Lacko, Marek; Bačenková, Darina; Amrichová, JuditaEditorial
Archivos de Medicina Universitaria
Materia
Autologous implantation Cartilage Cleanroom Chondrocytes Implantación autóloga Cartílago Sala blanca Condrocitos
Fecha
2016Referencia bibliográfica
José Manuel Muñoz Olmedo , Denisa Harvanová , Tímea Špaková , Ján Rosocha , Marek Lacko , Darina Bacenková and Judita Amrichová. Basics of isolation and cultivation of chondrocytes according to good laboratory practice. AMU. 2016; 4: 31-36
Resumen
OBJECTIVES: The objective of the present study was
to determine if chondrocytes isolated from human
cartilage of five elderly patients (mean age 63) with
osteoarthritis (stage 3) maintain their proliferation
and chondrogenic potential. Isolation and cultivation
of chondrocytes was performed according to
good laboratory practice (GLP) standards.
Methods: Chondrocytes were isolated from cartilage
biopsy by enzymatic digestion. Cultivation of cells was
performed in a controlled environment (cleanroom).
Phenotype characterization of chondrocytes
was achieved by flow cytometry analysis.
Results: Three weeks after cultivation, polygonal
structures typical of chondrocytes were observed,
but spindle/fibroblast like morphology was
also detected in cultured cells. Flow cytometric
analysis showed that chondrocytes were positive
for CD44 (98.35% ± 0.50), CD90 (97.15% ± 0.13) after
first passage (P1) and the cells were negative
for hematopoietic marker CD45 (0.21% ± 0.11).
ConclusionS: Human articular chondrocytes
obtained from five elderly patients with
osteoarthritis maintained a chondrocyte phenotype
and could be potentially used for autologous
implantation. We have standardized the conditions
for cultivation according to GLP standards to
minimize the risk of in vitro cell contamination.