Stromal cell-independent differentiation of human cord blood CD34+CD38− lymphohematopoietic progenitors toward B cell lineage

Yoshikawa, Yusuke; Hirayama, Fumiya; Kanai, Makoto I.; Ohkawara, Jun-ichi; Fujihara, Mitsuhiro; Miki, Yoshio; Sato, Norihiro; Kasai, Masaharu; Sekiguchi, Sadayoshi; Ikebuchi, Kenji

doi:10.1038/sj.leu.2401713

Cited by 6 publications

(5 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As is typical, from day 0–3, we observed a decreased level (from 92% to 22%) of CD34 antigens presenting on the surface of the fetal liver CD34+ cells (Figure 1A), resulting from the differentiation of cells due to the incubation with cytokines [39,40]. …”

Section: Resultsmentioning

confidence: 57%

“…Fetal liver CD34+ cells differentiate and loose the surface stem cell markers such as CD34 due to culture with cytokines [39,40]. Therefore, we confirmed the specificity of the CD34 (clone My10) antibody by testing the transduction efficiency of the 2.2 vector in the KG1a cell line, comprising an acute myelogenous leukemia cell line that expresses CD34 molecules, and stimulated PBMCs.…”

Section: Resultsmentioning

confidence: 61%

“…Targeting human CD34+ progenitor cells using VSV-G pseudotyped lentiviral vectors CD34+ cells were purified from fetal liver and cultured in IMDM with 20% FCS, IL-3, IL-6 and SCF [38]. As is typical, from day 0-3, we observed a decreased level (from 92% to 22%) of CD34 antigens presenting on the surface of the fetal liver CD34+ cells ( Figure 1A), resulting from the differentiation of cells due to the incubation with cytokines [39,40].…”

Section: Resultsmentioning

confidence: 63%

See 2 more Smart Citations

Targeted transduction of CD34+ hematopoietic progenitor cells in nonpurified human mobilized peripheral blood mononuclear cells

Liang¹,

Pariente²,

Morizono³

et al. 2009

The Journal of Gene Medicine

View full text Add to dashboard Cite

Background Conventional gene-therapy applications of hematopoietic stem cells (HSCs) involve purification of CD34+ progenitor cells from the mobilized peripheral blood, ex vivo transduction of the gene of interest into them, and reinfusion of the transduced CD34+ progenitor cells into patients. Eliminating the process of purification would save labor, time and money, while enhancing HSCs viability, transplantability and pluripotency. Lentiviral vectors have been widely used in gene therapy because they infect both dividing and nondividing cells and provide sustained transgene expression. One of the exceptions to this rule is quiescent primary lymphocytes, in which reverse transcription of viral DNA is not completed. Methods In the present study, we tested the possibility of targeting CD34+ progenitor cells within nonpurified human mobilized peripheral blood mononuclear cells (mPBMCs) utilizing vesicular stomatitis virus G (VSV-G) pseudotyped lentiviral vectors, based on the assumption that the CD34+ progenitor cells would be preferentially transduced. To further enhance the specificity of vector transduction, we also examined utilizing a modified Sindbis virus envelope (2.2) pseudotyped lentiviral vector, developed in our laboratory, that allows targeted transduction to specific cell receptors via antibody recognition. Results Both the VSV-G and 2.2 pseudotyped vectors achieved measurable results when they were used to target CD34+ progenitor cells in nonpurified mPBMCs. Conclusions Overall, the data obtained demonstrate the potential of ex vivo targeting of CD34+ progenitor cells without purification.

show abstract

Section: Resultsmentioning

confidence: 57%

Section: Resultsmentioning

confidence: 61%

Section: Resultsmentioning

confidence: 63%

See 1 more Smart Citation

Targeted transduction of CD34+ hematopoietic progenitor cells in nonpurified human mobilized peripheral blood mononuclear cells

Liang¹,

Pariente²,

Morizono³

et al. 2009

The Journal of Gene Medicine

View full text Add to dashboard Cite

show abstract

“…In another study, Bernhard et al (2000) reported that Hyper-IL6, a recombinant fusion protein of interleukin-6 with its specific receptor, promoted the differentiation of human hematopoietic stem cells (derived from either peripheral blood or bone marrow) into functional dendritic cells. In the case of umbilical cord blood, Flores-Guzmµn et al (2002) used a combination of recombinant cytokines to induce differentiation of the CD34+ enriched population into myeloid progenitors, while Yoshikawa et al (2000) reported differentiation into both myeloid and lymphoid lineages through the use of a recombinant cytokine cocktail. In addition to bone marrow and hematopoietic stem cells, the use of recombinant cytokines for the differentiation of hepatic stem cells (Wong and Tuan 1993) has also been reported.…”

Section: Development Of Defined Culture Milieu For Stem Cell Differenmentioning

confidence: 99%

An overview and synopsis of techniques for directing stem cell differentiation in vitro

Heng

Cao

Haider

et al. 2004

Cell and Tissue Research

View full text Add to dashboard Cite

The majority of studies on stem cell differentiation have so far been based in vivo, on live animal models. The usefulness of such models is limited, since it is much more technically challenging to conduct molecular studies and genetic manipulation on live animal models compared to in vitro cell culture. Hence, it is imperative that efficient protocols for directing stem cell differentiation into well-defined lineages in vitro are developed. The development of such protocols would also be useful for clinical therapy, since it is likely that the transplantation of differentiated stem cells would result in higher engraftment efficiency and enhanced clinical efficacy, compared to the transplantation of undifferentiated stem cells. The in vitro differentiation of stem cells, prior to transplantation in vivo, would also avoid spontaneous differentiation into undesired lineages at the transplantation site, as well as reduce the risk of teratoma formation, in the case of embryonic stem cells. Hence, this review critically examines the various strategies that could be employed to direct and control stem cell differentiation in vitro.

show abstract

“…4,8 Differentiation of B cells has also been reported for stromal cell–free cultures by use of a 2-step culture system or by adding supernatants from mesenchymal stromal cell cultures. 8,27 In another feeder cell–free in vitro system, CD34 + cells from cord blood or BM were cultured on plates coated with intercellular adhesion molecule-1–Fc in the presence of human IL-7, SCF, and FLT3L and subsequently maintained in cytokine-free medium. 28 …”

mentioning

confidence: 99%

Effects of autologous stromal cells and cytokines on differentiation of equine bone marrow–derived progenitor cells

Schwab¹,

Tallmadge²,

Matychak³

et al. 2017

ajvr

View full text Add to dashboard Cite

OBJECTIVE To develop an in vitro system for differentiation of equine B cells from bone marrow hematopoietic progenitor cells on the basis of protocols for other species. SAMPLE Bone marrow aspirates aseptically obtained from 12 research horses. PROCEDURES Equine bone marrow CD34+ cells were sorted by use of magnetic beads and cultured in medium supplemented with cytokines (recombinant human interleukin-7, equine interleukin-7, stem cell factor, and Fms-like tyrosine kinase-3), murine OP9 stromal cell preconditioned medium, and equine fetal bone marrow mesenchymal stromal cell preconditioned medium. Cells in culture were characterized by use of flow cytometry, immunocytofluorescence microscopy, and quantitative reverse-transcriptase PCR assay. RESULTS For these culture conditions, bone marrow–derived equine CD34+ cells differentiated into CD19+IgM+ B cells that expressed the signature transcription factors early B-cell factor and transcription factor 3. These conditions also supported the concomitant development of autologous stromal cells, and their presence was supportive of B-cell development. CONCLUSIONS AND CLINICAL RELEVANCE Equine B cells were generated from bone marrow aspirates by use of supportive culture conditions. In vitro generation of equine autologous B cells should be of use in studies on regulation of cell differentiation and therapeutic transplantation.

show abstract

Stromal cell-independent differentiation of human cord blood CD34+CD38− lymphohematopoietic progenitors toward B cell lineage

Abstract: To study the cytokine regulation of early stages of human Blymphopoiesis, we developed a stroma-free two-step culture system. Single human cord blood CD34؉ CD38

Cited by 6 publications

References 40 publications

Targeted transduction of CD34+ hematopoietic progenitor cells in nonpurified human mobilized peripheral blood mononuclear cells

Targeted transduction of CD34+ hematopoietic progenitor cells in nonpurified human mobilized peripheral blood mononuclear cells

An overview and synopsis of techniques for directing stem cell differentiation in vitro

Effects of autologous stromal cells and cytokines on differentiation of equine bone marrow–derived progenitor cells

Contact Info

Product

Resources

About