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

J Cell Biochem. 2011 Apr;112(4):1206-18. doi: 10.1002/jcb.23042.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / cytology
  • Adipose Tissue / metabolism
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / metabolism
  • CD146 Antigen / metabolism
  • Calcium-Binding Proteins
  • Cell Culture Techniques
  • Cell Differentiation*
  • Cell Proliferation*
  • Cell Separation
  • Cells, Cultured
  • Chondrocytes / cytology*
  • Cryopreservation
  • Female
  • Fetal Blood / cytology*
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Gene Expression
  • Humans
  • Immunophenotyping
  • Intercellular Signaling Peptides and Proteins / genetics
  • Male
  • Membrane Proteins / genetics
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism
  • Osteoclasts / cytology
  • Osteoclasts / metabolism
  • Ploidies
  • Reverse Transcriptase Polymerase Chain Reaction
  • Telomerase / metabolism
  • Telomere / genetics
  • Time Factors

Substances

  • CD146 Antigen
  • Calcium-Binding Proteins
  • DLK1 protein, human
  • Intercellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Telomerase