Hematopoietic Reconstitution of Syngeneic Mice with a Peripheral Blood-Derived, Monoclonal CD34-, Sca-1+, Thy-1low, c-kit+ Stem Cell Line

Lange, Claudia; Kaltz, Christine; Thalmeier, K; Hj, Kolb; Huss, Ralf

doi:10.1089/152581699320090

Cited by 47 publications

(43 citation statements)

References 28 publications

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“…Although we cannot completely rule out single donor cells below the detection limit, hematopoietic recovery in recipients is unlikely due to replacement with donor cells. This conclusion contradicts earlier results of hematopoietic recovery after myeloablative TBI with blood-derived mMSCs [11,15] showing donor characteristics in blood and BM. One fundamental difference between both cell sources is potential in vitro immortalization, altering BM seeding capability of MSC.…”

Section: Experimental Studiescontrasting

confidence: 99%

Mesenchymal stromal cells protect from consequences of HSCT-transplantation preparatory irradiation: insights into possible mechanisms

Lange¹,

Brunswig-Spickenheier²,

Reimer³

et al. 2016

CTT

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Ionizing irradiation is widely used as conditioning therapy in bone marrow (BM) transplantation. High-dose radiation treatment induces profound tissue damage, especially, of hematopoietic stem cells and progenitor cells. Efforts to improve clinical outcomes post-irradiation are focused on the hematopoietic stem cell niche. Mesenchymal stromal cells (MSCs) represent an integrative part of the BM stromal microenvironment. When co-transplanted with HSC, MSCs augment hematopoietic recovery after chemo-or radiotherapy. The aim of our study was to evaluate essential biological parameters of MSCs, with respect to their lineage-specific differentiation capacity, in vivo survival rates, as well as their ability to rescue lethally irradiated hosts. Materials and Methods. In vitro differentiation of human BM-derived MSCs (hMSCs) for hematopoietic (HSC) and endothelial cells (EC) was studied by reverse transcription-quantitative PCR (RT-qPCR) of lineage-specific surface markers and other proteins. To test in vivo ability of murine MSCs to rescue lethally irradiated (9.5 Gy) mice, the animals were transplanted with eGFP-marked murine MSCs (mMSCs). Long-term donor chimerism was assessed in blood, BM and thymus using CD45.2 and Y chromosome markers. A microarray analysis of bone marrow cells from MSC-transplanted animals was also performed, in order to compare their gene expression profiles to appropriate controls. ResultsUpon in vitro differentiation of hMSCs, the hematopoietically differentiated cells changed their gene expression towards a typical profile of progenitor and mature hematopoietic cells. A variety of transcription factors responsible for erythropoiesis, megakaryopoiesis, lympho-and myelopoiesis were up-regulated during differentiation in serum-containing media. A population of cells with small round or polymorphic nuclei was detected which expressed hematopoietic progenitor and mature antigen markers, albeit to a rather low degree. The same cells were able to acquire endothelial morphology and expressed endothelial genes upon cultivation with endothelial promoting factors. Following MSCs transplantation, the lethally irradiated mice showed normal hematopoietic recovery comparable to effects of HSC infusions. Seven months later, the recipients had normal distribution of peripheral blood cell populations. No evidence of donor chimerism was shown at any time CTT JOURNAL | VOLUME 5 | NUMBER 2 | JUNE 2016 51 EXPERIMENTAL STUDIES point posttransplant. The distribution kinetics of eGFP+ donor cells after i.v. transplantation identified fast disappearance from peripheral blood, reaching ca. 2% donor mMSC in peripheral blood after 8 hours and significant entrapment in lungs, however, without long-term persistence and embolization events. The microarray expression trial in BM cells from MSC-treated animals has shown upregulation of the genes which are beneficial to BM reconstitution, whereas the genes with supposed radiation-related BM deterioration were downregulated. Injection of MSC-derived microvesicles to le...

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Section: Experimental Studiescontrasting

confidence: 99%

Mesenchymal stromal cells protect from consequences of HSCT-transplantation preparatory irradiation: insights into possible mechanisms

Lange¹,

Brunswig-Spickenheier²,

Reimer³

et al. 2016

CTT

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“…There were some publications in recent years demonstrating that CD34-negative cells can also reconstitute the complete lympho-hematopoietic system of mice and sheep [18][19][20][21][22]. Although evidence is controversial , stromal cells as well as mesenchymal stem cells are considered to be CD34-negative [23].…”

Section: Discussionmentioning

confidence: 99%

Mesenchymal Differentiation and Organ Distribution of Established Human Stromal Cell Lines in NOD/SCID Mice

et al. 2001

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Two human stromal cell lines were established previously from bone marrow-derived primary long-term cultures by immortalization using the SV40 large T antigen and cellular cloning. After irradiation, the fibroblast-like cell lines L87/4 and L88/5 support hematopoietic differentiation of allogeneic cord blood cells in vitro. The stromal cells do not express CD34 and CD50, but some adhesion molecules and integrins, such as CD44, CD54 and CD58. Their expression profiles on RNA and protein levels are suggestive of their osteogenic potency. The quality and quantity of osteocalcin and osteopontin protein expression depended on the culture conditions. Expression of the osteogenic markers increased over time in culture, especially in cells growing in clusters. The stromal cells also expressed collagens I and V, but did not show any expression of collagens II and III. The potentially osteoblastic stromal cells were transplanted into NOD/ SCID recipient mice by intravenous injection and were found in various mesenchymal organs up to 10 weeks after transplantation. Osteocalcin-positive human stromal cells could be detected in the bone marrow, thymus, liver, brain and gut of the recipient animals. In summary, there is evidence that human bone-marrow-derived stromal cells have to be considered mesenchymal progenitors, persistently expressing osteogenic markers in vitro and in vivo.

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“…Long-term hematopoiesis may be dependent on a population of CD34-negative cells or some subset of CD34 + cells that we did not measure (ie true stem cells). 28,29 If this is true, the PBCD34 + cell count may better reflect this unmeasured cell population than the total CD34 + cell dose infused.…”

Section: Discussionmentioning

confidence: 99%

Predictive factors for long-term engraftment of autologous blood stem cells

Duggan

Guo

Luider

et al. 2000

Bone Marrow Transplant

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38− DR − ), peripheral blood CD34 + cell (PBCD34 + ) count on first apheresis day, or various clinical factors were associated with low blood counts 6 months post ASCT. Thirty-four patients were excluded from analysis either because of death (n = 17) or reinduction chemotherapy prior to 6 months post ASCT (n = 13), or because of lack of follow-up data (n = 4). Of the remaining 136 patients, 46% had low WBC (Ͻ4 × 10 9 /l), 41% low platelets (Ͻ150 × 10 9 /l), and 34% low hemoglobin (Ͻ120 g/l) at a median of 6 months following ASCT. By Spearman's rank correlation, both the total CD34 + cell dose/kg and the PBCD34 + count correlated with 6 month blood counts better than any subset of CD34 + cells or any clinical factor. The PBCD34 + count was overall a stronger predictor of 6 month blood counts than was the total CD34 + cells/kg infused. Both factors retained their significance in multivariate analysis, controlling for clinical factors. In conclusion, subsets of CD34 + cells and clinical factors are inferior to the total CD34 + cell dose/kg and PBCD34 + count in predicting 6 month blood counts following ASCT. Bone Marrow Transplantation (2000) 26, 1299-1304.

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Hematopoietic Reconstitution of Syngeneic Mice with a Peripheral Blood-Derived, Monoclonal CD34-, Sca-1+, Thy-1low, c-kit+ Stem Cell Line

Cited by 47 publications

References 28 publications

Mesenchymal stromal cells protect from consequences of HSCT-transplantation preparatory irradiation: insights into possible mechanisms

Mesenchymal stromal cells protect from consequences of HSCT-transplantation preparatory irradiation: insights into possible mechanisms

Mesenchymal Differentiation and Organ Distribution of Established Human Stromal Cell Lines in NOD/SCID Mice

Predictive factors for long-term engraftment of autologous blood stem cells

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