scholarly journals CH02 Peptide Promotes CD34+ Umbilical Cord Blood Hematopoietic Stem Cells Ex Vivo Expansion

2021 ◽  
Author(s):  
Yiqi Yang ◽  
Bihui Zhang ◽  
Junye Xie ◽  
Yuling Cai ◽  
Jia Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord Blood ◽  
Ex Vivo ◽  
Diabetic Mice ◽  
Serum Free Medium ◽  
Free Medium ◽  
Hematopoietic Stem ◽  
Serum Free ◽  
Cd34 Cells

Abstract Background: Umbilical cord blood (UCB) has been clinically used for human hematopoietic stem cells (HSCs) transplantation. However, limited numbers of the functional UCB-HSCs from single cord blood restricts its application in adults, while most of the strategies for stem cells expansion in vitro are either inefficient or costly. To overcome these obstacles, we evaluated the potential role of our newly identified CH02 peptide in ex vivo culture expansion of CD34+ UCB-HSCs. Methods: Enriched human CD34+ progenitor/stem cells populations were cultured in serum-free medium supplemented with different cytokines combinations for 8 days. These cytokines combinations included various concentration of CH02 peptide or the FLT3 ligand, with a cocktail of several growth factors such as IL-6, SCF and TPO. In addition, the global gene expression profile of the CD34+ cells cultured under different conditions were monitored through RNA-seq experiments. Furthermore, the expanded CD34+ cells were topically transplanted into the dorsal wounds of diabetic mice, and the wound closure was observed to evaluate the pro-repair ability of CH02-cultured CD34+ cells.Results: We herein report that the combination of CH02 peptide and other cytokines under the serum-free medium can effectively expand the CD34+ HSCs into 12-fold within 7 days while maintaining their stem cell properties. Moreover, CH02 peptide increased the anti-inflammatory and growth-promoting capacity of CD34+ cells, and thus accelerating wound healing of diabetic mice via promoting the anti-inflammatory and inhibiting the inflammatory factors.Conclusions: Together, our CH02 peptide demonstrated promising potentials to improve HSCs expansion for clinical application.

scholarly journals Ex Vivo Expansion and Transplantation of Hematopoietic Stem/Progenitor Cells Supported by Mesenchymal Stem Cells from Human Umbilical Cord Blood

2007 ◽  
Vol 16 (6) ◽  
pp. 579-585 ◽  
Author(s):  
Guo-Ping Huang ◽  
Zhi-Jun Pan ◽  
Bing-Bing Jia ◽  
Qiang Zheng ◽  
Chun-Gang Xie ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Cord Blood ◽  
Progenitor Cells ◽  
Umbilical Cord Blood ◽  
Ex Vivo ◽  
Hematopoietic Stem ◽  
Cd34 Cells

Human mesenchymal stem cells (MSCs) are multipotential and are detected in bone marrow (BM), adipose tissue, placenta, and umbilical cord blood (UCB). In this study, we examined the ability of UCB-derived MSCs (UCB-MSCs) to support ex vivo expansion of hematopoietic stem/progenitor cells (HSPCs) from UCB and the engraftment of expanded HSPCs in NOD/SCID mice. The result showed that UCB-MSCs supported the proliferation and differentiation of CD34+ cells in vitro. The number of expanded total nucleated cells (TNCs) in MSC-based culture was twofold higher than cultures without MSC (control cultures). UCB-MSCs increased the expansion capabilities of CD34+ cells, long-term culture-initiating cells (LTC-ICs), granulocyte-macrophage colony-forming cells (GM-CFCs), and high proliferative potential colony-forming cells (HPP-CFCs) compared to control cultures. The expanded HSPCs were transplanted into lethally irradiated NOD/SCID mice to assess the effects of expanded cells on hematopoietic recovery. The number of white blood cells (WBCs) in the peripheral blood of mice transplanted with expanded cells from both the MSC-based and control cultures returned to pretreatment levels at day 25 posttransplant and then decreased. The WBC levels returned to pretreatment levels again at days 45–55 posttransplant. The level of human CD45+ cell engraftment in primary recipients transplanted with expanded cells from the MSC-based cultures was significantly higher than recipients transplanted with cells from the control cultures. Serial transplantation demonstrated that the expanded cells could establish long-term engraftment of hematopoietic cells. UCB-MSCs similar to those derived from adult bone marrow may provide novel targets for cellular and gene therapy.

Characterization of Serum-Free Ex Vivo–Expanded Hematopoietic Stem Cells Derived from Human Umbilical Cord Blood CD133+Cells

2006 ◽  
Vol 15 (1) ◽  
pp. 70-78 ◽  
Author(s):  
Chao-Ling Yao ◽  
Yin-Hsun Feng ◽  
Xi-Zhang Lin ◽  
I-Ming Chu ◽  
Tzu-Bou Hsieh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Ex Vivo ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Hematopoietic Stem ◽  
Serum Free

Opposite Effects of M1 and M2 Macrophages on Hematopoietic Stem Cell Self-Renewal and Ex Vivo Expansion

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2909-2909
Author(s):  
Yi Luo ◽  
Lijian Shao ◽  
Jianhui Chang ◽  
Wei Feng ◽  
Chengcheng Li ◽  
...  
Keyword(s):  
Cord Blood ◽  
Ex Vivo ◽  
Serum Free Medium ◽  
Free Medium ◽  
Human Cord Blood ◽  
Self Renewal ◽  
Hematopoietic Stem ◽  
Expansion Medium ◽  
Cd34 Cells ◽  
Cord Blood Cd34

Abstract Macrophages (MΦ) are professional phagocytes in the innate immune system. They are not only involved in regulation of various immune functions and inflammation, but also exhibit plasticity in modulation of tissue regeneration and repair after being polarized into M1 and M2 MΦ by different inflammatory cytokines. In addition, several recent studies show that MΦ are a new constituent of the hematopoietic stem cells (HSCs) niche and play a role in regulation of HSCs maintenance and mobilization in bone marrow (BM). However, it is not known whether MΦ can regulate HSCs self-renewal and whether the effects of MΦ on HSCs can be influenced by differential MΦ polarization. This was investigated using an ex vivo HSCs expansion model consisting of mouse bone marrow LSK (Lin-sca-1+c-Kit+) cells cultured with or without MΦ in a mouse HSCs expansion medium (StemSpanTM serum-free medium supplemented with 20ng/ml of stem cell factor [SCF] and thrombopoietin [TPO]). We found that LSK cells were expanded about 20-, 15-, and 30-fold after 6 days of co-culture with MΦ harvested from mouse BM, spleen, and peritoneal cavity, respectively, whereas there was no significant expansion after culture without MΦ or with BM Gr-1high or Gr-1low monocytes. In addition, we found that M1-MΦ polarized by INFγ were more effective than IL4-polarized M2-MΦ in promoting LSK cells expansion ex vivo (45-fold vs. 15-fold). However, the promotion of LSK cells expansion by M1-MΦ resulted in about 88% reduction in HSCs as judged by 5-week cobblestone area forming cell (CAFC) assay. In contrast, M2-MΦ significantly promoted HSCs expansion. A greater expansion of HSCs was achieved after LSK cells were co-cultured with M2-MΦ for 9 days than for 6 days (20-fold vs. 6-fold). These findings suggest that M1-MΦ are more effective than M2-MΦ in promoting LSK cells or hematopoietic progenitor cells (HPCs) expansion, at the expense of HSCs self-renewal, whereas M2-MΦ can promote HPCs expansion as well as HSCs self-renewal. This suggestion is supported by results of serial transplantation and competitive repopulation unit (CRU) assays. CRU assay showed that LT-HSCs (e.g. 4-month CRU) were increased about 13 folds relative to the starting numbers of CRU in the input after LSK cells were co-cultured with M2-MΦ for 9 days, but were barely detectable after the cells were cultured without MΦ or with M1-MΦ. The inhibitory effect of M1-MΦ on HSCs self-renewal and expansion was attenuated by inhibition of inducible nitric oxide synthase (iNOS) activity with an inhibitor or knockout iNOS. Inhibition of arginase and/or cyclooxygenase activities with an inhibitor attenuated the promotion of HSCs self-renewal and expansion by M2-MΦ. More importantly, we found that human CD34+ cells, 8-week CAFC, and SCID mice repopulating cells (SRCs) were increased 42±14, 8±2.1, and 4 folds over the input values, respectively, after human cord blood CD34+ cells were co-cultured with M2-MΦ generated from human cord blood CD34- cells for 7 days in a human HSCs expansion medium (StemSpanTM serum-free medium supplemented with 50 ng/ml of SCF, TPO, and FLT-3 ligand). These findings demonstrate that M1-MΦ and M2-MΦ have opposite effects on HSCs self-renewal, which may be important for regulation of hematopoiesis under various pathological conditions in which MΦ are differentially polarized to M1 or M2 by diverse inflammatory cytokines. In addition, M2-MΦ may be used to promote human cord blood HSCs ex vivo expansion to make human cord blood transplantation available to more patients. Disclosures No relevant conflicts of interest to declare.

scholarly journals Unrestricted somatic stem cells from human umbilical cord blood grow in serum-free medium as spheres

2009 ◽  
Vol 9 (1) ◽  
pp. 101 ◽  
Author(s):  
Faten Zaibak ◽  
Paul Bello ◽  
Jennifer Kozlovski ◽  
Duncan Crombie ◽  
Haozhi Ang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Unrestricted Somatic Stem Cells

Transplantation of Ex Vivo Expanded Lin− Cells Selected from Autologous PBSC Grafts in Patients with Diagnosis of Lymphoma.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5198-5198
Author(s):  
Martin Klabusay ◽  
Zdenek Koristek ◽  
Jaroslava Vinklarkova ◽  
Jiri Mayer ◽  
Jiri Adler ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Ex Vivo ◽  
Immunomagnetic Separation ◽  
Serum Free Medium ◽  
Free Medium ◽  
Hematopoietic Stem ◽  
Serum Free ◽  
Cytokine Combination ◽  
Experimental Group

Abstract Background: Hematopoietic stem cells are able to regenerate hematopoiesis in all of its lineages. They are clinically used in transplantation of bone marrow or peripheral blood stem cells (PBSC) in patients with diagnosis of leukemia or lymphoma. While amount of hematopoietic stem cells is critical for the long-term engraftment, the amount of progenitor and precursor cells can influence time to engraftment, and it is critical for duration of neutropenia after transplantation. The methods of expansion of hematopoietic stem cells could reduce time to engraftment and decrease the risk of early post-transplant complications. Methods: Authors analyzed expansion of enriched hematopoietic stem cells (HSC), selected by immunomagnetic separation of Lin− cells from PBSC, in the culture of serum-free medium in vitro with all combinations of 5 cytokines (SCF, Flt-3-L, IL-3, IL-6, TPO) with and without G-CSF. Cell counts, morphology, immunophenotyping, and CFU-GM and CFU-Meg cultures were performed. Clinical transplantation protocol based on these results was developed. 10 patients with diagnosis of non-Hodgkin’s or Hodgkin’s lymphoma indicated for autologous transplantation, who signed the informed consent, were enrolled into the protocol. Except of standard PBSC graft, additional cells were collected, Lin− cells were selected by immunomagnetic separation, frozen and stored at Tissue bank. At day -14, Lin− cells were thawed and expanded in culture of serum-free medium with cytokines SCF, Flt-3-L, IL-3, IL-6 and G-CSF. Patients received high-dose chemotherapy regimen BEAM from day −7 to day −2. Progenitor cells expanded ex vivo in culture from Lin− cells were infused at day 0, following transplantation of PBSC. Patients were monitored, blood counts were performed, side effects were observed, and times to engraftment in granulocytes and platelets were calculated. Results: In experiments, the highest number of CFU-GM colonies was observed at day +14 with cytokine combination SCF+IL-3+Flt-3-L+ IL-6. The highest number of CFU-Meg colonies was observed in cytokine combination SCF+IL-3+TPO. G-CSF increased count and maturation of cells. Number of total cells grew 200 to 350 times at day +14. In the clinical protocol, 2 patients were excluded for technical reasons and 1 patient was excluded because of disease progression. 7 patients completed the protocol. The clinical procedure was free of serious adverse effects in all patients. Patients received doses from 5•107 to 3•109 cells. The group of patients was compared with historical controls - 142 patients treated at our department with autologous PBSC transplantation after BEAM regimen for diagnosis of lymphoma. Control group received higher average dose of CD34+ cells than experimental group: 7.7•106 / kg versus 6.6•106 / kg. Engraftment in granulocytes > 1000 / μl was shortened from 11 to 7.3 days in experimental group. Engraftment in platelets was not changed significantly (12 versus 11 days). Duration of neutropenia was shortened from 8 to 5.6 days. In patients, who received higher doses of infused cells > 2•109, average engraftment in granulocytes occurred at 6.3 days and duration of neutropenia was 5 days. Conclusions: HSC can be enriched from PBSC grafts, cultured and expanded ex vivo, and safely used in the cellular therapy protocols. The procedure resulted in significant shortening of critical period of neutropenia.

Hoechst dye efflux reveals a novel CD7+CD34− lymphoid progenitor in human umbilical cord blood

Blood ◽  
2000 ◽  
Vol 96 (6) ◽  
pp. 2125-2133 ◽  
Author(s):  
Robert W. Storms ◽  
Margaret A. Goodell ◽  
Alan Fisher ◽  
Richard C. Mulligan ◽  
Clay Smith
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Large Population ◽  
Lymphoid Cells ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Hematopoietic Stem ◽  
Cd34 Cells

Abstract A novel Hoechst 33342 dye efflux assay was recently developed that identifies a population of hematopoietic cells termed side population (SP) cells. In the bone marrow of multiple species, including mice and primates, the SP is composed primarily of CD34−cells, yet has many of the functional properties of hematopoietic stem cells (HSCs). This report characterizes SP cells from human umbilical cord blood (UCB). The SP in unfractionated UCB was enriched for CD34+ cells but also contained a large population of CD34− cells, many of which were mature lymphocytes. SP cells isolated from UCB that had been depleted of lineage-committed cells (Lin− UCB) contained CD34+ and CD34− cells in approximately equivalent proportions. Similar to previous descriptions of human HSCs, the CD34+Lin− SP cells were CD38dimHLA-DRdimThy-1dimCD45RA−CD71−and were enriched for myelo-erythroid precursors. In contrast, the CD34−Lin− SP cells were CD38−HLA-DR−Thy-1−CD71−and failed to generate myelo-erythroid progeny in vitro. The majority of these cells were CD7+CD11b+CD45RA+, as might be expected of early lymphoid cells, but did not express other lymphoid markers. The CD7+CD34−Lin− UCB SP cells did not proliferate in simple suspension cultures but did differentiate into natural killer cells when cultured on stroma with various cytokines. In conclusion, the human Lin− UCB SP contains both CD34+ multipotential stem cells and a novel CD7+CD34−Lin− lymphoid progenitor. This observation adds to the growing body of evidence that CD34− progenitors exist in humans.

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