title: Serum-free isolation and culture system to enhance the proliferation and bone regeneration of adipose tissue-derived mesenchymal stem cells.
abstract: Cell therapy using human mesenchymal stem cells (MSCs) is an attractive approach for many refractory diseases. Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are considered as a favorable tool due to its abundance in the body, easy proliferation, and high cytokine production potency. In order to avoid the risks associated with the use of fetal bovine serum (FBS) in culture that includes batch variations and contamination with pathogens, development of serum-free culture system has been initiated. We have formulated a completely serum-free culture medium (SFM) that could be used not only for the expansion of AT-MSCs but also for initial isolation. We demonstrate that the AT-MSCs isolated and cultured in serum-free medium (AT-MSCs/SFM) possess high proliferation capacity and differentiation potency to osteoblast, adipocyte, and chondrocyte lineages in vitro. In in vivo bone fraction model analysis, AT-MSCs/SFM showed higher bone repair potency and quality of the regenerated bone than the cells cultured in serum-containing medium (AT-MSCs/SCM). This was attributed to the (i) presence of translated cells in the bone, as evidenced by in vivo imaging of the illuminated translated cells and (ii) high level of expression and induction capacity of AT-MSCs/SFM for cytokine BMP2, CCL2, and CCL5. Taken together, we report a new serum-free culture system for AT-MSCs that is suitable for cell therapy.
authors.affiliation: Department of Regenerative Medicine and Stem Cell Biology, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305 8575, Japan, sato-k@cstimedia.com.
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Sato
authors.forename: Takehiro
authors.initials: T
authors.lastname: Itoh
authors.forename: Toshiki
authors.initials: T
authors.lastname: Kato
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kitamura
authors.forename: Sunil C
authors.initials: SC
authors.lastname: Kaul
authors.forename: Renu
authors.initials: R
authors.lastname: Wadhwa
authors.forename: Fujio
authors.initials: F
authors.lastname: Sato
authors.forename: Osamu
authors.initials: O
authors.lastname: Ohneda
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000704:Analysis of Variance; D001861:Bone Regeneration / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation / Q000502:physiology*; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D016895:Culture Media, Serum-Free; D016207:Cytokines / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007621:Karyotyping; D008297:Male; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D016895:Culture Media, Serum-Free; D016207:Cytokines; D017931:DNA Primers