title: Large-scale expansion of pre-isolated bone marrow mesenchymal stromal cells in serum-free conditions.
abstract: The regenerative potential of mesenchymal stromal or stem cells (MSCs) has generated tremendous interest for treating various degenerative diseases. Regulatory preference is to use a culture medium that is devoid of bovine components for stem cell expansion intended for therapeutic applications. However, a clear choice an alternative to fetal bovine serum (FBS) has not yet emerged. We have screened five different commercially available serum-free media (SFM) for their ability to support the growth and expansion of pre-isolated undifferentiated bone marrow-derived MSCs (BM-MSCs) and compared the results with cells grown in standard FBS-containing medium as control. In addition, based on initial screening results, BD Mosaic™ Mesenchymal Stem Cell Serum-free (BD-SFM) medium was evaluated in large-scale cultures for the performance and culture characteristics of BM-MSCs. Of the five different serum-free media, BD-SFM enhanced BM-MSCs growth and expansion in Cell STACK (CS), but the cell yield per CS-10 was less when compared to the control medium. The characteristics of MSCs were measured in terms of population doubling time (PDT), cell yield and expression of MSC-specific markers. Significant differences were observed between BD-SFM and control medium in terms of population doublings (PDs), cell yield, CFU-F and morphological features, whereas surface phenotype and differentiation potentials were comparable. The BD-SFM-cultured MSCs were also found to retain the differentiation potential, immune-privileged status and immunosuppressive properties inherent to MSCs. Our results suggest that BD-SFM supports large-scale expansion of BM-MSCs for therapeutic use.
authors.affiliation: Stempeutics Research Pvt. Ltd, Shirdi Sai Baba Cancer Hospital, Manipal, India.
authors.forename: Sanjay
authors.initials: S
authors.lastname: Gottipamula
authors.affiliation: Stempeutics Research Pvt. Ltd, Shirdi Sai Baba Cancer Hospital, Manipal, India.
authors.forename: Manjunatha S
authors.initials: MS
authors.lastname: Muttigi
authors.affiliation: Stempeutics Research Pvt. Ltd, Shirdi Sai Baba Cancer Hospital, Manipal, India.
authors.forename: S
authors.initials: S
authors.lastname: Chaansa
authors.affiliation: Stempeutics Research Pvt. Ltd, Shirdi Sai Baba Cancer Hospital, Manipal, India.
authors.forename: K M
authors.initials: KM
authors.lastname: Ashwin
authors.affiliation: Stempeutics Research Pvt. Ltd, Akshay Tech Park, Bangalore, India.
authors.forename: Nancy
authors.initials: N
authors.lastname: Priya
authors.affiliation: Stempeutics Research Pvt. Ltd, Akshay Tech Park, Bangalore, India.
authors.forename: Udaykumar
authors.initials: U
authors.lastname: Kolkundkar
authors.affiliation: Stempeutics Research Pvt. Ltd, Akshay Tech Park, Bangalore, India.
authors.forename: Swathi
authors.initials: S
authors.lastname: SundarRaj
authors.affiliation: Stempeutics Research Pvt. Ltd, Akshay Tech Park, Bangalore, India.
authors.forename: Anish Sen
authors.initials: AS
authors.lastname: Majumdar
authors.affiliation: Stempeutics Research Pvt. Ltd, Shirdi Sai Baba Cancer Hospital, Manipal, India.
authors.forename: Raviraja N
authors.initials: RN
authors.lastname: Seetharam
mesh_terms: D001681:Biological Assay; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D048430:Cell Shape; D003114:Colony-Forming Units Assay; D016895:Culture Media, Serum-Free; D006801:Humans; D056747:Immunomodulation; D016130:Immunophenotyping; D007165:Immunosuppression Therapy; D007700:Kinetics; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D055815:Young Adult
chemical_list: D016895:Culture Media, Serum-Free; D012710:Serum Albumin, Bovine
keywords: bone marrow; differentiation; large-scale expansion; mesenchymal stromal cells; phenotypic characterization; serum-free cultures