title: Impact of culture medium on CD4(+) CD25(high)CD127(lo/neg) Treg expansion for the purpose of clinical application.
abstract: A recently discovered population of lymphocytes, called T regulatory cells (Tregs), is characterized by expression of transcription factor Forkhead box P3 (FoxP3). These cells have been successfully used as therapeutic treatments and prophylaxis for graft-versus-host disease (GVHD) and diabetes and might become an attractive alternative to traditional immunotherapy. Here we evaluated how the type of culture medium and the type of serum can influence yield and quality of Tregs after in vitro expansion. We compared Treg fold of expansion and their phenotypical characteristics including expression of FoxP3, CD25, CD127, CD62L and CD45RA in three commercially available culture media (RPMI 1640 (Cellgro; Manassas VA, USA), SCGM (Cellgenix; Freiburg, Germany), and X-VIVO 20 (Lonza; Walkersville, MD, USA)) with addition of human serum (HS, 10%) or fetal bovine serum (FBS, 10%). Among the tested media, X-VIVO 20 supplemented with HS produced the highest yield after 17days of in vitro expansion (a median of 86-fold expansion, range 30-1365) and highest level of FoxP3 expression (a median of 66.8% of positive cells, range 56-84.8%) in CD4(+) CD25(hi)CD127(lo/neg) FACS sorted polyclonal Tregs. There was no difference in Tregs yield whether HS or FBS serum was used. In conclusion, the yield of the ex vivo expanded Tregs is related to the type of media applied. Supplementation of the culture with FBS or human serum is equally beneficial.
authors: Department of Surgery, Section of Transplantation, University of Chicago, IL, USA.
authors: Karolina
authors: K
authors: Gołąb
authors: Adam
authors: A
authors: Krzystyniak
authors: Natalia
authors: N
authors: Marek-Trzonkowska
authors: Ryosuke
authors: R
authors: Misawa
authors: Ling Jia
authors: LJ
authors: Wang
authors: Xiaojun
authors: X
authors: Wang
authors: Olivia
authors: O
authors: Cochet
authors: Martin
authors: M
authors: Tibudan
authors: Paulina
authors: P
authors: Langa
authors: J Michael
authors: JM
authors: Millis
authors: Piotr
authors: P
authors: Trzonkowski
authors: Piotr
authors: P
authors: Witkowski
mesh_terms: D015703:Antigens, CD / Q000276:immunology; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D051858:Forkhead Transcription Factors / Q000276:immunology; D006801:Humans; D044967:Serum / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology
chemical_list: D015703:Antigens, CD; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors