<h3>Background</h3> Cell therapy, which involves the transplantation of stem cells, progenitor cells, primary cells and/or genetically modified, may facilitate the repair of damaged tissue and organs. In orthopedics, the use of chondrocytes to treat articular cartilage lesions is already present in clinical practice. However, there is a demand for the standardization of cells isolation, characterization and expansion. The aim of this study was to conduct an integrative literature review about the methods used, purposing at the development of a standardized research laboratory protocol. <h3>Methods</h3> Bardin's method (2011) was used to characterize in electronic spreadsheets the findings referring to the descriptors "Chondrocyte", "Chondrocyte isolation", "Chondrocyte cultivation" and "Chondrocyte characterization" on the ClinicalTrials, PubMed and Scopus platforms. Based on the methodological parameters, the efficiency of the techniques found in the literature review, a new protocol was developed for the isolation, characterization and expansion of chondrocytes obtained from the knee cartilage. <h3>Results</h3> The Scopus platform presented more studies available (62.3%). The chi-squared test showed statistical significance for the descriptor "Chondrocyte" in the number of results retrieved (p= 0.004) and useful (p=0.04). A total of 78.9% of the surgeries were performed in the knee region, mainly for the treatment of articular cartilage injuries (57.4%). The review identified few studies about the objective, even so it was possible to establish the proposed protocol. The knee cartilage was the region with the highest cells for isolation (31.4%) which we standardized in our study to 3 mm² biopsy dissociated enzymatically with type I collagenase. Expansion time was detailed in 30.8% of the studies, with two weeks being standardized for confluence at passage three. Chondrocyte characterization was not very explained or performed in the researched articles, however we established a protocol by flow cytometry with the following markers: anti-CD14, anti-CD45, anti-CD19, anti-CD44, anti-CD73, anti-CD90, anti-CD151, anti-CD49C, anti-HLA-DR, anti-CD34, anti-CD105, anti-CD29. <h3>Conclusion</h3> The integrative literature review was sufficient to demonstrate studies about the proposed aim, allowing the construction of a research protocol of isolation, expansion and characterization ready to be applied in laboratory research.