CD133 Marks a Stem Cell Population That Drives Human Primary Myelofibrosis

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1601-1601
Author(s):  
Ioanna Triviai ◽  
Thomas Stuebig ◽  
Birte Niebuhr ◽  
Kais Hussein ◽  
Asterios Tsiftsoglou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Population ◽  
Myeloid Cell ◽  
Primary Myelofibrosis ◽  
Jak2v617f Mutation ◽  
Stem Cell Population ◽  
Cell Populations ◽  
Cd34 Cells

Abstract Primary Myelofibrosis (PMF) is a chronic myeloproliferative neoplasm of alleged stem cell origin. To define the characteristics of malignant PMF stem cells previous studies have focused on the isolation and xenotransplantation of circulating and/or splenic, PMF patient - derived CD34+ stem/progenitor cells. Despite the reported engraftment of CD34+ cell pool, former analyses failed to reproduce major PMF parameters attributed to abnormal human myeloid cell differentiation. The focus of our work was to identify the stem cell population responsible for initiation and development of PMF. To assess the presence of malignant stem cells we analyzed peripheral blood of 30 PMF patients for expression of LT-HSC antigen CD133. To exclude committed myeloid and lymphoid circulating progenitors we performed lineage depletion of PBMCs and isolated CD133+ and/or CD34+ stem cells. Variable CD133+/CD34 ± and CD133-/CD34+ stem cell fractions from 15 PMF patients were assessed for their clonogenic potential in semisolid media and for reproduction of PMF morbidity in a xenotransplantation mouse model. JAK2V617F mutation was used as a genetic marker to track clonal evolution both in vitro and in vivo. In patients' PBMC we detected the consistent presence of a CD133+ population ranging from 0.3% to >30%, which varies in the expression of CD34. CD133 marks overlapping but also distinct cell populations as compared to CD34. To determine the differentiation potential of disparate stem cell populations, CD133+CD34+, CD133-CD34+ and CD133+CD34- cells were subfractionated from PB of 7 patients and assessed for clonogenic capacity. Strikingly, CD133+CD34+ cells exhibited multipotent, bipotent, and unipotent myeloid (including erythroid) and endothelial-like output, whereas CD133-CD34+ cells gave rise predominantly to lineage-restricted granulocyte/monocyte (GM) progenitors or endothelial-like progenitors. Thus, in contrast to circulating CD133-/CD34+ cells in PMF patients, CD133+ cells have a broader and more robust differentiation capacity to all myeloid cell types, including megakaryocyte /erythrocyte lineages. Four JAK2V617F+ patient samples were used to assess mutation burden at the single-cell level from representative colony types. Obtained results demonstrate an early acquisition of JAK2V617F mutation in the primitive CD133+ stem cell compartments, but also revealed an unexpected variability in the genotypes of emerging progenitors. Homozygous JAK2617F/617F progenitors were detectable in all analyzed patient samples, even if a relative low JAK2V617F burden (30%) was determined from the initial pool of CD133+ cells. A disproportionately high incidence of a homozygous JAK2V617F genotype was observed in erythroid progenitors, indicating a skewing for this lineage. Homozygosity was additionally detected in megakaryocytic and multipotent progenitors. In vivo xenotransplantation experiments of various subfractions confirm the origin of multipotent JAK2V617F+ progenitors from CD133+/CD34± stem cells. Transplantation of PMF patient-derived CD133+/CD34± stem cells in immuno-compromised mice induces abnormal human JAK2V617F+ erythroid, megakaryocytic, and monocytic differentiation, splenomegaly, bone marrow/splenic fibrosis and anemia, reproducing many aspects of PMF development. Our data provide the first evidence for the existence of a CD133+ LT-HSC population responsible for development of PMF. It is for the first time demonstrated that JAK2V617F mutation in PMF occurs at the level of a multipotent stem cell, from which all abnormal myeloid cells emanate during evolution of the disease. Identification of the stem cell compartment involved in the triggering and progression of PMF provides the basis to elucidate the nature of the complex niche interactions in myeloproliferative neoplasms. Disclosures: No relevant conflicts of interest to declare.

Detection of a Stem Cell Population with Hemangioblastic Characteristics in Primary Myelofibrosis (PMF),

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3860-3860
Author(s):  
Ioanna N Trivai ◽  
Thomas Stuebig ◽  
Anita Badbaran ◽  
Ursula Gehling ◽  
Asterios Tsiftsoglou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Stem Cell ◽  
Peripheral Blood ◽  
Primary Myelofibrosis ◽  
Jak2v617f Mutation ◽  
Cell Subpopulation ◽  
Stem Cell Population ◽  
Allele Burden

Abstract Abstract 3860 Primary myelofibrosis (PMF) comprises a myeloproliferative neoplasia accompanied by imbalance of various tissues of the mesoderm, let alone the hematopoietic tissue. Involvement of multiple hematopoietic lineages during disease progression suggests the clonality of myelofibrosis that can be attributed to an initial stem cell defect at the very early stage of the stem cell hierarchy. Hematopoietic and endothelial phenotypes of circulating multipotent stem cells in patient peripheral blood, along with the increased microvascular density in the bone marrow, leads to the hypothesis that the critical event in PMF involves malignant transformation of a stem cell with hemangioblastic potential. Former studies have provided functional evidence that activated JAK2 signalling in primitive human hematopoietic cells is sufficient to drive key processes involved in the pathogenesis of the disease. In this study, the functionality and differentiation potential of circulating primitive JAK2V617F+ stem cells from primary myelofibrosis patients is assessed. Primitive stem cells were isolated from peripheral blood of 25 patients. All patients participating in the study were diagnosed with primary myelofibrosis, have been untreated, and were found positive for JAK2V617F mutation. Isolated stem cells were analysed for purity and assessed for the expression of markers characteristic for the hemangioblast phenotype (CD34, CD133, CD45, VEGFR2, VE-Cadherin, E-Cadherin, CD31) with flow cytometry. Genomic DNA was isolated from various stem cell populations to determine the mutational status by PCR. Our results indicate that long term repopulating stem cells circulating in peripheral blood bear the JAK2V617F mutation. Hemangioblast resembling populations within the isolated prime stem cells were also found positive for the mutation. Long term repopulating stem cells bearing different allele burden for JAK2V617F mutation from PMF patient peripheral blood were expanded for up to 4 months. Various colonies formed after seeding in semisolid media were characterised by morphological features (CFU-GEMM, CFU-GM, CFU-E, CFU-M, CFU-Endo) and expressing genes by quantitative PCR. Moreover, allele burden determination for various progenitors of both hematopoietic and endothelial lineages was performed. JAK2V617F allele burden varied within individual progeny phenotypes, indicating the acquisition of the mutation that boosts the outgrowth of the malignant clone within the hemangioblast compartment of the bone marrow. Endothelial and macrophage progenitors appear heterozygotic while all rest progenitors of various hematopoietic lineages can be either heterozygotic or homozygotic. This indicates high genomic instability of the JAK2V617F+ malignant clone as it is driven into hematopoietic differentiation. Our results indicate the existence of a malignant clone with hemangioblast phenotype in PMF which can differentiate into hematopoietic and/or endothelial progenitors in vitro. Our experiments shed light to the pathogenesis of PMF by characterising the potential of the defective stem cell subpopulation responsible for the disease. Disclosures: No relevant conflicts of interest to declare.

Differential Expression of SLAM Family Receptor Markers in Normal Human Hematopoietic Stem Cells and Their Malignant Counterpart in MDS and AML.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1897-1897
Author(s):  
Ramon V. Tiu ◽  
Jennifer J. Powers ◽  
Abdo Haddad ◽  
Ying Jiang ◽  
Jaroslaw P. Maciejewski
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Stem Cell ◽  
Hematopoietic Stem Cells ◽  
Cell Population ◽  
Stem Cell Population ◽  
Leukemic Stem Cell ◽  
Hematopoietic Stem ◽  
Cd34 Cells ◽  
Slam Family

Abstract Members of the signaling lymphocytic activation molecule (SLAM) family, including CD150, CD48 and CD244, were shown to precisely distinguish more committed lineage restricted progenitor cells from pluripotent and multipotent murine hematopoietic stem cells (HSC; Kiel et al; 2005 Cell). Similar SLAM profiles may also be present on HSC subsets in humans. We hypothesized that these SLAM markers may be indicators not only of stem cell potential in normal hematopoiesis but also distinguish a subset of the most immature malignant precursors of leukemia. In agreement with the concept of a “cancer stem cell,” the presence of leukemic stem cell population may be an indicator of important clinical and biological properties. We first tested the distribution of CD150, CD48 and CD244 antigens on human CD34+ cells derived from 7 control individuals using 4-color flow cytometry. CD34+ cells were measured in the blast gate based on side scatter and CD45 expression. Within CD34+ blasts, expression of CD48, CD150, and CD244 was detected on 16.71%±9.69, 6.53%±2.93, and 26.92%±6.95 of cells respectively. Subsequently, we investigated SLAM expression in 9 immature leukemic cell lines, including KG-1, K562, U937, HEL, HL60, MKN-95, NB-4, Kasumi and UT7, and found increased expression of SLAM markers in KG-1 (CD48+, CD150+, CD244+) and Kasumi (CD48−, CD150−, CD244+). Consequently, none of the leukemic cells showed pluripotent/multipotent SLAM profiles. We then compared the SLAM marker expression on blasts from patients with AML and MDS with that of CD34+ cells from normal controls. We studied a total of 28 patients: 11 MDS (2 low grade, 5 advanced MDS, 3 MDS/MPD overlap) and 10 AML (FAB: 3 M1, 2 M2, 1 M3, 2 M4/M4E0 and 2 M6). In our cohort, 8/10 AML patients expressed one of the three SLAM markers; 6/10 were CD150−CD48−CD244+ (63.57%±6.96) and 2/10 were CD150+CD48−CD244−(46%±10.96) suggestive of the presence of either pluripotent or multipotent leukemic stem cell phenotype. In the MDS cohort, 8/11 patients expressed one of three SLAM markers, 7/11 were CD150−CD48−CD244+ (41.21% ± 8.9) and 1/11 were CD150+CD48−CD244− (1.26%±0.59) again consistent with a profile derived from either pluripotent or multipotent stem cells. None of the MDS and AML patients had either co-expression of CD244 and CD48 or increased expression of CD48 alone. Two of the M1 type AML patients with CD150−CD48−CD244+ phenotype received prior chemotherapy and achieved complete remission on bone marrow biopsy and flow cytometry using traditional blast markers. In some, we conclude that the SLAM receptor markers may be associated with certain types of leukemic blasts and may be useful in the identification of leukemic stem cell population in both MDS and AML.

The Leukemic Stem Cell in Polycythemia Vera and Primary Myelofibrosis Is Distinct From the Initiating JAK2 V617F-Positive Hematopoiertic Stem Cell

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 613-613
Author(s):  
Jonathan M. Gerber ◽  
Brownhilda Ngwang ◽  
Hao Zhang ◽  
Donna M. Williams ◽  
Ophelia Rogers ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Population ◽  
Jak2 V617f ◽  
Primary Myelofibrosis ◽  
Cell Populations ◽  
Aldh Activity ◽  
Hematopoietic Progenitor ◽  
Allele Burden ◽  
Cd34 Cells ◽  
5Q Deletion

Abstract Abstract 613 The chronic myeloproliferative disorders (MPD), polycythemia vera (PV) and primary myelofibrosis (PMF) are clonal disorders involving a multipotent hematopoietic stem cell (HSC) but the identity of the involved HSC is a matter of controversy. For example, involvement of lymphoid cells in the MPD clone as determined by JAK2 V617F expression implies that these disorders arise in a pluripotent HSC. However, evidence has been presented that PV CD34+ HSC expressing JAK2 V617F are predisposed to erythroid differentiation, while mathematical modeling of JAK2 V617F-positive MPD suggests that these disorders arise in a hematopoietic progenitor cell that acquires JAK2 V617F, and then a mutation conferring self-renewal. Recently, a high level of aldehyde dehydrogenase (ALDHhigh) activity was used to distinguish normal CD34+CD38− HSC from committed hematopoietic progenitor cells, which lack ALDH activity, and also to separate leukemic stem cells (LSC) from CD34+/CD38−(ALDHhigh) HSC due to the presence of lower ALDH activity(ALDHint) in the LSC. We, therefore, sought to determine whether ALDH activity could be used to define the HSC involved in JAK2 V617F-positive PV and PMF, and whether leukemic transformation in these disorders was associated with a change in ALDH activity in the involved stem cell. To this end, we studied the immunophenotypic characteristics and ALDH activity of circulating CD34+ cells from 6 PV, 6 PMF (3 PMF and 3 post PV/MF) and 3 post MPD acute myelogenous leukemia (AML) patients (2 PV and 1PMF). CD34+ cells were isolated from peripheral blood using immunomagnetic bead technology with a purity of 95 % and a viability of 98 %, and analyzed for CD34 and CD38 expression and ALDH activity by flow cytometry. Cell sorting was carried out for measurement of the JAK2 V617F allele burden in discrete cell populations using purified genomic DNA and either a quantitative allele-specific assay or pyrosequencing. Where informative, sorted cell populations were analyzed by FISH for chromosomal abnormalities. As a per cent of total leukocytes, the median CD34+ cell fraction was 0.07 (range 0.01–0.2) for PV; 0.5 (range 0.4–1.3) for PV/MF and 2.3 (range 0.2–2.9) for PMF. The CD34+CD38− fraction was 17.8% of total PV CD34+ cells, 20.7% of total PV/MF CD34+ cells (p = 0.69) and 51.6 % of total PMF CD34+ cells (p= 0.003 and p= 0.007 respectively), indicating greater expansion of the PMF CD34+CD38− cell population, even though there was not a significant difference in disease duration between the three groups. ALDH activity was high in the CD34+CD38− cell population of all three groups. In 9/9 patients studied, the JAK2 V617F mutation was present in the CD34+CD38−(ALDHhigh) cell population at approximately 80 % of the patients' neutrophil JAK2 V617F allele burden, substantiating that the JAK2 V617F mutation was present in primitive PV and PMF HSC. We next analyzed the CD34+CD38− cell population for ALDH activity in 1 PMF and 2 PV patients who had transformed to AML. In addition to an CD34+/CD38−(ALDHhigh) cell population, all three patients had an additional CD34+/CD38− population with lower ALDH activity (ALDHint), identical to that of LSC. Importantly, this latter CD34+/CD38−(ALDHint) cell population was not present before AML transformation in any patient. In 2 of 2 patients studied, the CD34+/CD38−(ALDHint) cell population expressed JAK2 V617F with an allele burden comparable to the CD34+/CD38−(ALDHhigh) population. Significantly, in one of the patients, who had acquired a 5q- deletion, the chromosomal abnormality was present only in the CD34+CD38−(ALDHint) cell population. In conclusion, these data support the contention that in PV and PMF, JAK2 V617F is acquired in a primitive CD34+CD38−(ALDHhigh) HSC. In addition, in contrast to PV and PV/MF, expansion of total CD34+ cells in PMF was also accompanied by differential expansion of this primitive CD34+CD38−(ALDHhigh) population. Furthermore, AML transformation in PV and PMF appeared to occur in an LSC, which had an aberrant ALDH activity pattern (ALDHint) identical to that seen in de novo AML LSC. The 5q- deletion, a molecular marker characteristic of AML, also tracked with the LSC cell population but not with the CD34+CD38−(ALDHhigh) cell population, while JAK2 V617F was equivalently expressed by both cell populations. This observation supports the contention that with respect to AML transformation in PV and PMF, JAK2 V617F is essentially a passenger lesion. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Genotype-restricted growth and aging patterns in hematopoietic stem cell populations of allophenic mice.

1990 ◽  
Vol 171 (5) ◽  
pp. 1547-1565 ◽  
Author(s):  
G Van Zant ◽  
B P Holland ◽  
P W Eldridge ◽  
J J Chen
Keyword(s):  
Stem Cells ◽  
Cell Proliferation ◽  
Stem Cell ◽  
Blood Cell ◽  
Cell Population ◽  
Stromal Cell ◽  
Stem Cell Population ◽  
Cell Populations ◽  
Stem Cell Proliferation ◽  
Stem Cell Populations

We have studied contributions to hematopoiesis of genetically distinct stem cell populations in allophenic mice. Chimeras were made by aggregating embryos of inbred strains known to differ with respect to stem cell population kinetics. One partner strain (DBA/2) has previously been shown to normally have a stem cell (CFU-S) population of which 24% are in S-phase of the cell cycle, whereas the homologous population of the other partner strain (C57BL/6) was characterized by having only 2.6% in cycle (7). Contributions of the chimeric stem cell population to mature blood cell pools were studied throughout the life of the mice and intrinsic differences in stem cell function and aging were reflected in dynamic patterns of blood cell composition. The DBA/2 stem cell population was eclipsed by stem cells of the C57BL/6 genotype and, after 1.5-3 yr, the hemato-lymphoid composition of 22 of 27 mice studied for this long had shifted by at least 25 percentage points toward the C57BL/6 genotype. 8 of the 27 had hematolymphoid populations solely of C57BL/6 origin. To test whether or not a population of stem cells with an inherently higher cycling rate (DBA/2) might have a competitive advantage during repopulation, we engrafted allophenic marrow into lethally irradiated (C57BL/6 x DBA/2)F1 recipients. DBA/2 hematopoiesis was predominant early, far outstripping its representation in the marrow graft. Perhaps as a consequence of inherently greater DBA/2 stem cell proliferation, the populations of developmentally more restricted precursor populations (CFU-E, BFU-E, CFU-GM, CFU-GEMM) showed an overwhelming DBA/2 bias in the first 2-3 mo after engraftment. However, as in the allophenic mice themselves during the aging process, the C57BL/6 genotypic representation was ascendant over the subsequent months. The shift toward C57BL/6 genotype was first documented in the marrow and spleen precursor cell populations and was subsequently reflected in the circulating, mature blood cells. Bone marrow-derived stromal cell cultures from engrafted mice were studied and genotypic analyses showed donor representation in stromal cell populations that reflected donor hematopoietic contributions in the same recipient. Results from these studies involving two in vivo settings (allophenic mice and engraftment by allophenic marrow) are consistent with the notion that a cell autonomous difference in stem cell proliferation confers on one population a competitive repopulating advantage, but at the expense of longevity.

scholarly journals Identification of a human adult cardiac stem cell population with neural crest origin

2019 ◽  
Author(s):  
Anna Höving ◽  
Madlen Merten ◽  
Kazuko Elena Schmidt ◽  
Isabel Faust ◽  
Lucia Mercedes Ruiz-Perera ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Neural Crest ◽  
Cell Population ◽  
Stem Cell Population ◽  
Cell Populations ◽  
Cardiac Stem Cell ◽  
Adult Human ◽  
Neural Crest Origin ◽  
Stem Cell Populations

ABSTRACTCardiovascular diseases are the major cause of death worldwide, emphasizing the necessity to better understand adult human cardiac cell biology and development. Although the adult heart was considered as a terminally differentiated organ, rare populations of cardiac stem cells (CSCs) have been described so far, with their developmental origin and endogenous function still being a matter of debate.Here, we identified a Nestin+/S100+/CD105+/Sca1+/cKit-population of CSCs in the myocardium of the adult human heart auricle. Isolated cells showed expression of characteristic neural crest-derived stem cell (NCSC) markers and kept their genetic stability during cultivationin vitro. Cultivated hCSCs efficiently gave rise to functional, beating cardiomyocytes, osteoblasts, adipocytes and neurons. Global transcriptome analysis via RNAseq showed a high similarity between the expression profiles of Nestin+/S100+/CD105+/Sca1+/cKit-hCSCs and adult human NCSCs from the nasal cavity (inferior turbinate stem cells, ITSCs). In detail, 88.1 % of all genes were significantly expressed in both stem cell populations particularly including common NCSC-markers. Based on these observations, we suggest a similar developmental origin of both stem cell populations.In summary, we identified a human adult cardiac stem cell population with neural crest-origin, which may also contribute to endogenous cardiac tissue homeostasis and tissue repairin vivo.

scholarly journals Sarcoma Stem Cells: Do We Know What We Are Looking for?

Sarcoma ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Matteo Trucco ◽  
David Loeb
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cancer Stem Cells ◽  
Malignant Transformation ◽  
Cell Population ◽  
Mesenchymal Cells ◽  
Heterogeneous Group ◽  
Stem Cell Population ◽  
Cell Populations ◽  
Stem Cell Populations

Sarcomas represent a heterogeneous group of cancers thought to originate from malignant transformation of mesenchymal cells. There is increasing evidence that many, if not all, sarcomas contain within them tumor-initiating, or “cancer stem,” cells responsible for the initiation, maintenance, and potentially relapse and metastasis of the tumor. Various techniques have been adopted in recent years to identify putative sarcoma stem cell populations. The goal of this paper is to summarize the criteria used to identify a stem cell population, describe the more prominent markers and techniques used to isolate cancer stem cells in sarcomas, and review the evidence for the existence of cancer stem cells in sarcomas.

Growth Factor Deprivation Combined with Prolonged Inhibition of BCR-ABL Does Not Eradicate Functional CML Stem Cells

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4222-4222
Author(s):  
Ashley Hamilton ◽  
Gudmundur V Helgason ◽  
Elaine Allan ◽  
Michael J. Alcorn ◽  
Franck E Nicolini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Growth Factors ◽  
Growth Factor ◽  
Cell Population ◽  
Continuous Treatment ◽  
Stem Cell Population ◽  
Stem Cell Markers ◽  
Cd34 Cells ◽  
Growth Factor Deprivation

Abstract Chronic myeloid leukaemia (CML) is a myeloproliferative disorder that originates in a haemopoietic stem cell (HSC) and is characterised by the BCR-ABL oncogene. We have previously shown that cancer stem cells exist in all patients with newly diagnosed CML. These cells are primitive, quiescent, transplantable in NOD/SCID mice and insensitive to the tyrosine kinase inhibitor (TKI) imatinib. This intrinsic resistance may in part be explained by high expression of BCR-ABL in the most primitive cells. Our overall aim was to achieve complete and prolonged inhibition of BCR-ABL within CML CD34+ cells in vitro, using the highest clinically achievable concentration of a novel dual Src/ABL TKI, dasatinib, in serum free medium in the absence of growth factors. The drug was replenished every 3 days and cells remaining at 12 days were analysed. Despite growth factor deprivation combined with 12 days continuous treatment with dasatinib 150nM, 10% of the starting CD34+ cells were recovered. CFSE stained CML cells showed that treatment with dasatinib resulted in a “backing up” of cells within the undivided cell population and earlier cell divisions, demonstrating that dasatinib, like imatinib, is anti-proliferative. Inhibition of P-CrkL and hence BCR-ABL was maximal at the earliest time-point (day 4) and in the cells that were able to proliferate through several divisions, likely the most mature cells. Cells which remained undivided/quiescent and maintained expression of stem cell markers, CD34 and CD133, demonstrated significantly elevated levels of P-CrkL, as compared to the mature cycling populations. Following drug washout, the remaining viable cells achieved 5-fold expansion when cultured in growth factors for 7 days. Moreover, these cells were able to form colonies following culture in LTCIC assays, which were BCR-ABL positive by FISH and did not show gene mutation. Retrospective studies which assessed the efficacy of TKI treatment times and concentrations also showed that transient dasatinib exposure for 1 hour at 150nM, resulted in a ~80% reduction of viable CML cells when assessed 72 hours later and was as effective as 72 hours continuous exposure at concentrations as high as 1000nM. Our next approach will be to combine BCR-ABL knock-down using shRNA with dasatinib to fully inhibit BCR-ABL in the stem cell population in an effort to determine whether these cells are dependent on BCR-ABL for survival and proliferation.

Stem cells in mammary health and milk production

2021 ◽  
Vol 16 ◽  
Author(s):  
Ratan K Choudhary ◽  
Fenq-Qi Zhao
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Cell Population ◽  
Adult Stem Cells ◽  
Stem Cell Population ◽  
Mammary Stem ◽  
In Vitro Expansion

: Adult stem cells like mammary and mesenchymal stem cells have received significant attention because these stem cells (SCs) possess therapeutic potential in treating many animal diseases. These cells can be administered in an autologous or allogenic fashion, either freshly isolated from the donor tissue or previously cultured and expanded in vitro. Expansion of adult stem cells is a prerequisite before therapeutic application because sufficient numbers are required in dosage calculation. Stem cells directly and indirectly (by secreting various growth factors and angiogenic factors called secretome) act to repair and regenerate injured tissues. Recent studies on mammary stem cells showed in vivo and in vitro expansion ability by removing the blockage of asymmetrical cell division. Compounds like purine analogs (xanthosine, xanthine, and inosine) or hormones (progesterone and bST) help increase stem cell population by promoting cell division. Such methodology of enhancing stem cells number, either in vivo or in vitro, may help in preclinical studies for translational research like treating diseases like mastitis. The application of mesenchymal stem cells has also been shown to benefit mammary gland health due to the ‘homing’ property of stem cells. In addition to that, the multiple positive effects of stem cell secretome are on mammary tissue healing and killing bacteria is novel in the production of quality milk. This systematic review discusses some of the studies on stem cells that have been useful in increasing the stem cell population and increasing mammary stem/progenitor cells. Finally, we provide insights into how enhancing mammary stem cell population could potentially increase terminally differentiated cells, ultimately leading to more milk production.

scholarly journals 3D Differentiation Of Mammosphere Derived Macaca fascicularis’s Mammary Stem Cells

2019 ◽  
Vol 3 (1) ◽  
Author(s):  
Silmi Mariya
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Mammary Gland ◽  
Cell Population ◽  
Mammary Stem Cell ◽  
Stem Cell Population ◽  
Stem Cell Markers ◽  
Mammary Stem Cells ◽  
Surgical Biopsy ◽  
Mammary Stem

The mammary gland contains adult stem cells that are capable of self-renewal.  This population plays an important role in the development of mammary gland and breast cancer pathogenesis. The studies of mammary stem cells are limited due to the difficulty to acquire and expand adult stem cell population in an undifferentiated state. In this study, we developed mammosphere cultures of nulliparous cynomolgus monkeys (Macaca fascicularis; Mf) as a culture system to enrich mammary stem cells. This species has similarity of mammary gland structure as humans including anatomy, developmental stages, and lobule profile of mammary gland. The use of stem cells from primate animals is essential to bridge the knowledge gaps resulting from stem cell research using rodents for clinical trials in human. Small samples of mammary tissues were collected by surgical biopsy; cells were cultured as monolayer and cryopreserved. Cryopreserved cells were cultured into mammospheres, and the expression of markers for mammary stem cells was evaluated using qPCR. Cells were further differentiated with 3D approaches to evaluate morphology and organoid budding. The study showed that mammosphere culture resulted in an increase in the expression of mammary stem cell markers with each passage. The 3D differentiation in matrigel allowed for organoid formation. Mammary gland stem cells have been successfully differentiated which characterized by CSN2 marker expression and differentiation regulators marker STAT5 and GATA3. The results indicate that mammospheres can be successfully developed derived from breast tissue of nulliparous Mf collected via surgical biopsy. As the mammosphere allows for enrichment of mammary stem cell population, the findings also suggest that a 3-dimensional system is efficient as in-vitro model to study mammary stem cells and a useful system to study mammary differentiation in regards to cancer prevention.

scholarly journals PIWIL1 Drives Chemoresistance in Multiple Myeloma by Modulating Mitophagy and the Myeloma Stem Cell Population

2022 ◽  
Vol 11 ◽  
Author(s):  
Yajun Wang ◽  
Lan Yao ◽  
Yao Teng ◽  
Hua Yin ◽  
Qiuling Wu
Keyword(s):  
Multiple Myeloma ◽  
Drug Resistance ◽  
Stem Cell ◽  
Cell Population ◽  
Side Population ◽  
Chemotherapeutic Agents ◽  
Stem Cell Population ◽  
Exact Function

As an important member of the Argonaute protein family, PIWI-like protein 1 (PIWIL1) plays a key role in tumor cell viability. However, the exact function of PIWIL1 in multiple myeloma (MM) and the underlying mechanism remain unclear. Here, we revealed that PIWIL1 was highly expressed in myeloma cell lines and newly diagnosed MM patients, and that its expression was notably higher in refractory/relapsed MM patients. PIWIL1 promoted the proliferation of MM cells and conferred resistance to chemotherapeutic agents both in vitro and in vivo. More importantly, PIWIL1 enhanced the formation of autophagosomes, especially mitophagosomes, by disrupting mitochondrial calcium signaling and modulating mitophagy-related canonical PINK1/Parkin pathway protein components. Mitophagy/autophagy inhibitors overcome PIWIL1-induced chemoresistance. In addition, PIWIL1 overexpression increased the proportion of side population (SP) cells and upregulated the expression of the stem cell-associated genes Nanog, OCT4, and SOX2, while its inhibition resulted in opposite effects. Taken together, our findings demonstrated that PIWIL1 induced drug resistance by activating mitophagy and regulating the MM stem cell population. PIWIL1 depletion significantly overcame drug resistance and could be used as a novel therapeutic target for reversing resistance in MM patients.

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