Human stem-progenitor cells from neonatal cord blood have greater hematopoietic expansion capacity than those from mobilized adult blood.

OBJECTIVE In this study we compared the hematopoietic capacity of CD34+ cell preparations from neonatal cord blood (CB) vs adult mobilized peripheral blood (PBSC) before and after ex vivo culture. METHODS CD34+ cell preparations purified from CB or PBSC were cultured in serum-free medium containing FKT: FLT-3 ligand (FL), KIT ligand (KL), and thrombopoietin (TPO). RESULTS After 1-4 weeks ex vivo culture, CB CD34+ cell preparations had greatly increased numbers of total cells, CD34+ cells, and colony-forming cells (CFC). In contrast, ex vivo-cultured PBSC CD34+ cell preparations generated far less in vitro assessed hematopoietic capacity. Nonobese diabetic severe combined immunodeficient mouse (NOD/SCID) engrafting potential (SEP) was maintained in ex vivo-cultured CB CD34+ cell preparations, whereas ex vivo-cultured PBSC lost SEP. CB CD34+ cells continued to proliferate throughout 3 weeks ex vivo, whereas after 1 week, no additional cell divisions were detected in PBSC CD34+ cells. After 3 weeks in culture, the average CB CD34+ cell had divided more than 5 times, as compared to only 2 times for the average PBSC CD34+ cell. CONCLUSION CB CD34+ cell preparations generated massively increased in vitro assessed hematopoietic capacity and maintained SEP during 1- to 4-week ex vivo cultures. In contrast, ex vivo-cultured PBSC CD34+ cell preparations generated far less in vitro assessed hematopoietic capacity and decreased SEP. The differences in the in vitro proliferative indices of membrane dye-labeled CD34+ cells from CB vs PBSC correlated with these functional differences.

[1]  C. Civin,et al.  Quantity and quality of engrafting cells in cord blood and autologous mobilized peripheral blood. , 1999, Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation.

[2]  C. Civin,et al.  Measurement of CD34+ cells in bone marrow by flow cytometry. , 1993, Journal of hematotherapy.

[3]  R. Henschler,et al.  Hematopoietic recovery of ex vivo perfusion culture expanded bone marrow and unexpanded peripheral blood progenitors after myeloablative chemotherapy , 2001, Bone Marrow Transplantation.

[4]  C. Civin,et al.  Ex vivo culture of cord blood CD34+ cells expands progenitor cell numbers, preserves engraftment capacity in nonobese diabetic/severe combined immunodeficient mice, and enhances retroviral transduction efficiency. , 1999, Human gene therapy.

[5]  Lesley J. Murray,et al.  Thrombopoietin, kit ligand, and flk2/flt3 ligand together induce increased numbers of primitive hematopoietic progenitors from human CD34+Thy-1+Lin- cells with preserved ability to engraft SCID-hu bone. , 1998, Blood.

[6]  L. Naldini,et al.  Stable transduction of quiescent CD34(+)CD38(-) human hematopoietic cells by HIV-1-based lentiviral vectors. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[7]  P. Rebulla,et al.  Placental/umbilical cord blood transplantation. , 1999, Haematologica.

[8]  C. Eaves,et al.  Direct evidence for multiple self-renewal divisions of human in vivo repopulating hematopoietic cells in short-term culture. , 1999, Blood.

[9]  C. Eaves,et al.  Rapid differentiation of a rare subset of adult human lin(-)CD34(-)CD38(-) cells stimulated by multiple growth factors in vitro. , 1999, Blood.

[10]  Mamoru Ito,et al.  Expansion of human NOD/SCID-repopulating cells by stem cell factor, Flk2/Flt3 ligand, thrombopoietin, IL-6, and soluble IL-6 receptor. , 2000, The Journal of clinical investigation.

[11]  David A. Williams,et al.  Nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mouse as a model system to study the engraftment and mobilization of human peripheral blood stem cells. , 1998, Blood.

[12]  D. Sutherland,et al.  The ISHAGE guidelines for CD34+ cell determination by flow cytometry. International Society of Hematotherapy and Graft Engineering. , 1996, Journal of hematotherapy.

[13]  S. Richardson,et al.  Mixtures of distributions: inference and estimation , 1995 .

[14]  E. A. Parent.  MIXTURES OF DISTRIBUTIONS , 1962 .

[15]  M. Fackler,et al.  Human CD34+ cell preparations contain over 100-fold greater NOD/SCID mouse engrafting capacity than do CD34- cell preparations. , 2001, Experimental hematology.

[16]  J. Dick,et al.  Expansion of human cord blood CD34(+)CD38(-) cells in ex vivo culture during retroviral transduction without a corresponding increase in SCID repopulating cell (SRC) frequency: dissociation of SRC phenotype and function. , 2000, Blood.

[17]  A. Pecora.  Progress in clinical application of use of progenitor cells expanded with hematopoietic growth factors , 2001, Current opinion in hematology.

[18]  I McNiece,et al.  Ex vivo expansion of hematopoietic progenitor cells and mature cells. , 2001, Experimental hematology.

[19]  K. Cornetta,et al.  Ex vivo expansion of hematopoietic stem and progenitor cells: are we there yet? , 1999, Journal of hematotherapy.

[20]  G. Wagemaker,et al.  In vivo expansion of hemopoietic stem cells , 1998, Stem cells.

[21]  P. Schirmacher,et al.  Interleukin-6 and the soluble interleukin-6 receptor induce stem cell factor and Flt-3L expression in vivo and in vitro. , 2001, Experimental hematology.

[22]  M. Nieda,et al.  Clinical application of in vitro expansion of cord blood. , 1998, Bone marrow transplantation.

[23]  A. Nagler,et al.  Interleukin-6 receptor-interleukin-6 fusion proteins with enhanced interleukin-6 type pleiotropic activities. , 1997, European cytokine network.

[24]  K. Ando,et al.  Ex vivo generation of CD34(+) cells from CD34(-) hematopoietic cells. , 1999, Blood.

[25]  G. de Haan,et al.  Alternatives to stem cell renewal from a developmental viewpoint. , 1997, Experimental hematology.

[26]  C. Civin,et al.  Mature human hematopoietic cells in donor bone marrow complicate interpretation of stem/progenitor cell assays in xenogeneic hematopoietic chimeras. , 1998, Experimental hematology.

[27]  M. Ogawa,et al.  Human bone marrow CD34- cells engraft in vivo and undergo multilineage expression that includes giving rise to CD34+ cells. , 1998, Experimental hematology.

[28]  M. Baccarani,et al.  Phenotypic characterization of immunomagnetically purified umbilical cord blood CD34+ cells. , 1999, Blood cells, molecules & diseases.

[29]  W. Piacibello,et al.  Engraftment in nonobese diabetic severe combined immunodeficient mice of human CD34(+) cord blood cells after ex vivo expansion: evidence for the amplification and self-renewal of repopulating stem cells. , 1999, Blood.

[30]  C. Eaves,et al.  Efficient retrovirus-mediated gene transfer to transplantable human bone marrow cells in the absence of fibronectin. , 2000, Blood.

[31]  C. Civin,et al.  In vivo engraftment potential of clinical hematopoietic grafts. , 1998, Journal of investigative medicine : the official publication of the American Federation for Clinical Research.

[32]  A. Zander,et al.  Comparison of outcomes of unrelated bone marrow and umbilical cord blood transplants in children with acute leukemia. , 2001, Blood.

[33]  J. Wagner,et al.  Survival after transplantation of unrelated donor umbilical cord blood is comparable to that of human leukocyte antigen-matched unrelated donor bone marrow: results of a matched-pair analysis. , 2001, Blood.

[34]  W. Piacibello,et al.  Extensive amplification and self-renewal of human primitive hematopoietic stem cells from cord blood. , 1997, Blood.

[35]  J. Adamson,et al.  Processing and cryopreservation of placental/umbilical cord blood for unrelated bone marrow reconstitution. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[36]  K. Ando,et al.  Murine Stromal Cell Line HESS‐5 Maintains Reconstituting Ability of Ex Vivo‐Generated Hematopoietic Stem Cells from Human Bone Marrow and Cytokine‐Mobilized Peripheral Blood , 2000, Stem Cells.

[37]  M. Roncarolo,et al.  Ex vivo manipulations alter the reconstitution potential of mobilized human CD34+ peripheral blood progenitors , 1999, Leukemia.

[38]  Sylvia Richardson,et al.  Markov Chain Monte Carlo in Practice , 1997 .

[39]  I. Fichtner,et al.  Differential Kinetics of Primitive Hematopoietic Cells Assayed In Vitro and In Vivo During Serum‐Free Suspension Culture of CD34+ Blood Progenitor Cells , 1999, Stem cells.

[40]  H. W. Lee,et al.  Dissociation between stem cell phenotype and NOD/SCID repopulating activity in human peripheral blood CD34(+) cells after ex vivo expansion. , 2001, Experimental hematology.

[41]  A. Nagler,et al.  The soluble interleukin-6 (IL-6) receptor/IL-6 fusion protein enhances in vitro maintenance and proliferation of human CD34(+)CD38(-/low) cells capable of repopulating severe combined immunodeficiency mice. , 1999, Blood.

[42]  C. von Kalle,et al.  Previously undetected human hematopoietic cell populations with short-term repopulating activity selectively engraft NOD/SCID-beta2 microglobulin-null mice. , 2001, The Journal of clinical investigation.

[43]  L. Qiu,et al.  Ex vivo expansion of CD34+ umbilical cord blood cells in a defined serum-free medium (QBSF-60) with early effect cytokines. , 1999, Journal of hematotherapy & stem cell research.

[44]  G. Gilmore,et al.  Ex vivo expansion of human umbilical cord blood and peripheral blood CD34(+) hematopoietic stem cells. , 2000, Experimental hematology.

[45]  J. Dick,et al.  Quantitative Analysis Reveals Expansion of Human Hematopoietic Repopulating Cells After Short-term Ex Vivo Culture , 1997, The Journal of experimental medicine.