Rotating three‐dimensional dynamic culture of adult human bone marrow‐derived cells for tissue engineering of hyaline cartilage

Abstract

The method of constructing cartilage tissue from bone marrow-derived cells in vitro is considered a valuable technique for hyaline cartilage regenerative medicine. Using a rotating wall vessel (RWV) bioreactor developed in a NASA space experiment, we attempted to efficiently construct hyaline cartilage tissue from human bone marrow-derived cells without using a scaffold. Bone marrow aspirates were obtained from the iliac crest of nine patients during orthopedic operation. After their proliferation in monolayer culture, the adherent cells were cultured in the RWV bioreactor with chondrogenic medium for 2 weeks. Cells from the same source were cultured in pellet culture as controls. Histological and immunohistological evaluations (collagen type I and II) and quantification of glycosaminoglycan were performed on formed tissues and compared. The engineered constructs obtained using the RWV bioreactor showed strong features of hyaline cartilage in terms of their morphology as determined by histological and immunohistological evaluations. The glycosaminoglycan contents per mg DNA of the tissues were 10.01 AE 3.49 mg/mg DNA in the case of the RWV bioreactor and 6.27 AE 3.41 mg/mg DNA in the case of the pellet culture, and their difference was significant. The RWV bioreactor could provide an excellent environment for three-dimensional cartilage tissue architecture that can promote the chondrogenic differentiation of adult human bone marrow-derived cells. ß

Transient vascularization of transplanted human adult–derived progenitors promotes self-organizing cartilage

“…Subcutaneous transplantation was also used in a part of this study aimed at future clinical use, since the primary indication for elastic cartilage regeneration will be craniofacial deformities, as noted in our previous article (9). As controls, CPCs from the same donor were also subjected to conventional pellet culture (28). Approximately 2 × 10 6 cells were placed in a 15-ml polypropylene tube and centrifuged at 500 g for 5 minutes at 4°C.…”

Usefulness of Agarose Mold as a Storage Container for Three-Dimensional Tissue-Engineered Cartilage

“…It was reported that the large-sized tissue-engineered constructs were cultured, using roating wall vessel (RWV) [17]. When the RWV was used to culture the mesenchymal stem cells, favorable results of cartilage regeneration have been found [18]. However, the floating culture requires specific devices, a problem remains with regard to transport, and, basically, only rounded shapes, such as spherical and circular shapes, can be used because mechanical forces can be spread out.…”

A flow perfusion bioreactor with controlled mechanical stimulation: Application in cartilage tissue engineering and beyond

“…Ohyabu and colleagues [30] used cultured rabbit BMSCs without a scaffold, and obtained cartilaginous constructs with higher expression of GAG and Col II compared to pellets. Human BMSCs cultured in an RWV without a scaffold, resulted in statistically signi icantly higher concentrations of GAG and Col II compared to pellets [31]. Yoshioka and colleagues [32] reported successful regeneration of cartilage in osteochondral-defective Japanese white rabbits using cartilaginous constructs obtained from culturing BMSCs.…”