title: Isolation and characterization of mesenchymal stem cells from human umbilical cord blood: reevaluation of critical factors for successful isolation and high ability to proliferate and differentiate to chondrocytes as compared to mesenchymal stem cells from bone marrow and adipose tissue.
abstract: Human umbilical cord blood (CB) is a potential source for mesenchymal stem cells (MSC) capable of forming specific tissues, for example, bone, cartilage, or muscle. However, difficulty isolating MSC from CB (CB-MSC) has impeded their clinical application. Using more than 450 CB units donated to two public CB banks, we found that successful cell recovery fits a hyper-exponential function of time since birth with very high fidelity. Additionally, significant improvement in the isolation of CB-MSC was achieved by selecting cord blood units having a volume ≥90  ml and time ≤2  h after donor's birth. This resulted in 90% success in isolation of CB-MSC by density gradient purification and without a requirement for immunoaffinity methods as previously reported. Using MSC isolated from bone marrow (BM-MSC) and adipose tissue (AT-MSC) as reference controls, we observed that CB-MSC exhibited a higher proliferation rate and expanded to the order of the 1 × 10(9)  cells required for cell therapies. CB-MSC showed karyotype stability after prolonged expansion. Functionally, CB-MSC could be more readily induced to differentiate into chondrocytes than could BM-MSC and AT-MSC. CB-MSC showed immunosuppressive activity equal to that of BM-MSC and AT-MSC. Collectively, our data indicate that viable CB-MSC could be obtained consistently and that CB should be reconsidered as a practical source of MSC for cell therapy and regenerative medicine using the well established CB banking system.
authors.affiliation: Cell Therapy Research and Development Laboratory, New York Blood Center, New York, New York 10065, USA.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Zhang
authors.forename: Masako
authors.initials: M
authors.lastname: Hirai
authors.forename: Susana
authors.initials: S
authors.lastname: Cantero
authors.forename: Rodica
authors.initials: R
authors.lastname: Ciubotariu
authors.forename: Ludy
authors.initials: L
authors.lastname: Dobrila
authors.forename: Allen
authors.initials: A
authors.lastname: Hirsh
authors.forename: Koichi
authors.initials: K
authors.lastname: Igura
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Satoh
authors.forename: Izuru
authors.initials: I
authors.lastname: Yokomi
authors.forename: Toshihide
authors.initials: T
authors.lastname: Nishimura
authors.forename: Satoru
authors.initials: S
authors.lastname: Yamaguchi
authors.forename: Kotaro
authors.initials: K
authors.lastname: Yoshimura
authors.forename: Pablo
authors.initials: P
authors.lastname: Rubinstein
authors.forename: Tsuneo A
authors.initials: TA
authors.lastname: Takahashi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D051929:CD146 Antigen / Q000378:metabolism; D002135:Calcium-Binding Proteins; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D015925:Cryopreservation; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D016130:Immunophenotyping; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics; D008297:Male; D008565:Membrane Proteins / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D010010:Osteoclasts / Q000166:cytology / Q000378:metabolism; D011003:Ploidies; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019098:Telomerase / Q000378:metabolism; D016615:Telomere / Q000235:genetics; D013997:Time Factors
chemical_list: D051929:CD146 Antigen; D002135:Calcium-Binding Proteins; C512941:DLK1 protein, human; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D019098:Telomerase