Pub Date : 2016-12-20DOI: 10.15406/JSRT.2016.01.00045
S. Bartelmez, C. Storey, P. Iversen, F. Ruscetti
HSC transplantation efficiency can be improved by modulating cell surface molecules such as CD26 and CXCR4.1,2 This is important in cases where donor cell numbers are limiting, as in cord blood stem cell transplantation in adults where high morbidity and slow engraftment occur3 due to limiting numbers of CD34+ cells and possibly more differentiate progeny.4 The approach involving ex vivo expansion of primitive HSC to increase the total transplantable cell number has not been clinically useful.5,6 Here, we took the approach of enhancing HSC engraftment without expansion by transiently inhibiting endogenous TGFβ1 expression in HSC using minimal ex vivo manipulation. We previously reported that TGFβ1 directly and reversibly inhibits the initial cell divisions of murine LTR-HSC as well hematopoietic progenitor cells in vitro.7,8 TGFβ1 has been shown to be a primary regulator of LTR-HSC quiescence (G0) in bone marrow niches.9 Adult mice with a conditional knock-out of the TGFβ1 receptor exhibited essentially normal in vivo hematopoiesis.10 However, HSC/progenitor cells released from G0 quiescence in vitro by TGFβ neutralizing antibodies resulted in improved retroviral gene transfer.11,12 Furthermore, inhibiting Smad signaling, the intracellular regulators of TGFβ1 signaling, promoted HSC self-renewal in vivo.13 Here we studied highly enriched murine LTRHSC from lineagenegative bone marrow cells followed by FACS sequential selection of low retention of the fluorescent, viable dyes Höechst 33342 (Hö) (binding A-T base pairs of DNA) and Rhodamine 123 (Rh) (binding predominately activated mitochondria membranes). Low retention of the dyes is due to both high efflux rates and low target binding. Our previous studies showed that selecting cells that are lin-, Hö low (~G0),14 c-kit+7,15 identifies both LTRHSC and short -term repopulating (STR-HSC). A further selection step based on Rh fluorescence resolves the LTR-HSC (low Rh) and STR-HSC (high Rh). LTR-HSC are predominantly quiescent9 compared to STR-HSC, and are unique in their long-term repopulating ability due a high probability of selfreplication while at the same time generating large numbers of STRHSC16 Untreated LTR-HSC have poor short-term repopulating ability early after transplant, however over time in vivo give rise to daughter cells that possess efficient short-term repopulating ability. We show that transient inhibition of endogenous TGFβ1 in LTR-HSC ex vivo
{"title":"Transient Inhibition of Endogenous Transforming Growth Factor-β1 (Tgfβ1) in Hematopoietic Stem Cells Accelerates Engraftment and Enhances Multi-Lineage Repopulating Efficiency","authors":"S. Bartelmez, C. Storey, P. Iversen, F. Ruscetti","doi":"10.15406/JSRT.2016.01.00045","DOIUrl":"https://doi.org/10.15406/JSRT.2016.01.00045","url":null,"abstract":"HSC transplantation efficiency can be improved by modulating cell surface molecules such as CD26 and CXCR4.1,2 This is important in cases where donor cell numbers are limiting, as in cord blood stem cell transplantation in adults where high morbidity and slow engraftment occur3 due to limiting numbers of CD34+ cells and possibly more differentiate progeny.4 The approach involving ex vivo expansion of primitive HSC to increase the total transplantable cell number has not been clinically useful.5,6 Here, we took the approach of enhancing HSC engraftment without expansion by transiently inhibiting endogenous TGFβ1 expression in HSC using minimal ex vivo manipulation. We previously reported that TGFβ1 directly and reversibly inhibits the initial cell divisions of murine LTR-HSC as well hematopoietic progenitor cells in vitro.7,8 TGFβ1 has been shown to be a primary regulator of LTR-HSC quiescence (G0) in bone marrow niches.9 Adult mice with a conditional knock-out of the TGFβ1 receptor exhibited essentially normal in vivo hematopoiesis.10 However, HSC/progenitor cells released from G0 quiescence in vitro by TGFβ neutralizing antibodies resulted in improved retroviral gene transfer.11,12 Furthermore, inhibiting Smad signaling, the intracellular regulators of TGFβ1 signaling, promoted HSC self-renewal in vivo.13 Here we studied highly enriched murine LTRHSC from lineagenegative bone marrow cells followed by FACS sequential selection of low retention of the fluorescent, viable dyes Höechst 33342 (Hö) (binding A-T base pairs of DNA) and Rhodamine 123 (Rh) (binding predominately activated mitochondria membranes). Low retention of the dyes is due to both high efflux rates and low target binding. Our previous studies showed that selecting cells that are lin-, Hö low (~G0),14 c-kit+7,15 identifies both LTRHSC and short -term repopulating (STR-HSC). A further selection step based on Rh fluorescence resolves the LTR-HSC (low Rh) and STR-HSC (high Rh). LTR-HSC are predominantly quiescent9 compared to STR-HSC, and are unique in their long-term repopulating ability due a high probability of selfreplication while at the same time generating large numbers of STRHSC16 Untreated LTR-HSC have poor short-term repopulating ability early after transplant, however over time in vivo give rise to daughter cells that possess efficient short-term repopulating ability. We show that transient inhibition of endogenous TGFβ1 in LTR-HSC ex vivo","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"67 1","pages":"1-10"},"PeriodicalIF":0.0,"publicationDate":"2016-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79538572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-12-02DOI: 10.15406/JSRT.2016.01.00044
E. Sitnicka, C. Storey, S. Bartelmez
Several studies have demonstrated that the hematopoietic stem cell (HSC) compartment consists of long-term repopulating (LTR) and short-term repopulating (STR) HSC. Here we describe an improved purification approach that identifies both LTR- and STR- HSC as being lineage negative, c-kit positive, with very low Hoechst 33342 retention (Lin-, c-kit+, Holow). However, further selection of cells based on their differential retention of Rhodamine 123 resolves cells into LTR-HSC and STR-HSC. We show that our sort method highly enriches for LTR-HSC (Rhlow) and STR-HSC (Rhhigh), and demonstrate that the Rhlow cells as single transplanted cells are able to engraft 70% of mice in a competitive long-term repopulating assay. We then describe several in vitro assays that resolve Rhlow and Rhhigh cells based on the ability of single cells to survive, form clones, vary the time to their first cell division, express a high proliferative potential (HPP) or to generate HPP daughter cells at the 2- to 8-cell stage. In the presence of IL-3 alone, single Rhlow cells divided rarely and then formed only small clones (~8 cells). In contrast, Rhhigh readily divided in IL-3 alone and went on to form large clones (~10,000 cells). However, in the presence of IL-3+IL-6+SCF, both cell populations cloned in vitro with high efficiency (>90%), although the proportion of HPP clones was significantly higher in the Rhlow cell fraction (~90% vs ~40%). Furthermore, In addition, we show that the time required by Rhlow cells to undergo their first cell division in vitro is relatively non-synchronous and longer than that of Rhhigh cells. In addition, an analysis of daughter cells generated during the initial cell divsions of Rhlow or Rhhigh cells in vitro showed that expansion or maintenance of total HPP daughter cells occurred only in the Rhlow cell fraction. We measured the proliferative potential of daughter cells derived from single Rhlow and Rhhigh cells at the 2-8 cell stage. At the 2-cell stage, Rhlow cells generated an increased number of HPP daughter cells (↑1.4-fold) compared to Rhhigh cells that appeared to maintain the total number of HPP daughter cells (1.0-fold). However, by the 8-cell stage, the total number of HPP daughter cells generated by Rhlow cells expanded to nearly double that of starting HPP numbers (↑1.9 fold), compared to a decline in total HPP daughter cells in 8-cell Rhhigh clones (↓0.5 fold). Our studies at the 2-cell stage directly demonstrate symmetrical divisions (2 HPP per 2 daughter cells) that result in HPP expansion. Thus, these studies of growth factor responsivness of purified HSC (survival, cloning efficiency, time to the first cell division) and differentiation pathways (their ability to generate HPP daughter cells) identify means to differentiate LTR- and STR HSC in vitro.
{"title":"Functional resolution of long-term and short-term-hematopoietic stem cells","authors":"E. Sitnicka, C. Storey, S. Bartelmez","doi":"10.15406/JSRT.2016.01.00044","DOIUrl":"https://doi.org/10.15406/JSRT.2016.01.00044","url":null,"abstract":"Several studies have demonstrated that the hematopoietic stem cell (HSC) compartment consists of long-term repopulating (LTR) and short-term repopulating (STR) HSC. Here we describe an improved purification approach that identifies both LTR- and STR- HSC as being lineage negative, c-kit positive, with very low Hoechst 33342 retention (Lin-, c-kit+, Holow). However, further selection of cells based on their differential retention of Rhodamine 123 resolves cells into LTR-HSC and STR-HSC. We show that our sort method highly enriches for LTR-HSC (Rhlow) and STR-HSC (Rhhigh), and demonstrate that the Rhlow cells as single transplanted cells are able to engraft 70% of mice in a competitive long-term repopulating assay. We then describe several in vitro assays that resolve Rhlow and Rhhigh cells based on the ability of single cells to survive, form clones, vary the time to their first cell division, express a high proliferative potential (HPP) or to generate HPP daughter cells at the 2- to 8-cell stage. In the presence of IL-3 alone, single Rhlow cells divided rarely and then formed only small clones (~8 cells). In contrast, Rhhigh readily divided in IL-3 alone and went on to form large clones (~10,000 cells). However, in the presence of IL-3+IL-6+SCF, both cell populations cloned in vitro with high efficiency (>90%), although the proportion of HPP clones was significantly higher in the Rhlow cell fraction (~90% vs ~40%). Furthermore, In addition, we show that the time required by Rhlow cells to undergo their first cell division in vitro is relatively non-synchronous and longer than that of Rhhigh cells. In addition, an analysis of daughter cells generated during the initial cell divsions of Rhlow or Rhhigh cells in vitro showed that expansion or maintenance of total HPP daughter cells occurred only in the Rhlow cell fraction. We measured the proliferative potential of daughter cells derived from single Rhlow and Rhhigh cells at the 2-8 cell stage. At the 2-cell stage, Rhlow cells generated an increased number of HPP daughter cells (↑1.4-fold) compared to Rhhigh cells that appeared to maintain the total number of HPP daughter cells (1.0-fold). However, by the 8-cell stage, the total number of HPP daughter cells generated by Rhlow cells expanded to nearly double that of starting HPP numbers (↑1.9 fold), compared to a decline in total HPP daughter cells in 8-cell Rhhigh clones (↓0.5 fold). Our studies at the 2-cell stage directly demonstrate symmetrical divisions (2 HPP per 2 daughter cells) that result in HPP expansion. Thus, these studies of growth factor responsivness of purified HSC (survival, cloning efficiency, time to the first cell division) and differentiation pathways (their ability to generate HPP daughter cells) identify means to differentiate LTR- and STR HSC in vitro.","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"21 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91166146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-11-30DOI: 10.15406/JSRT.2016.01.00043
F. Khallaf, E. Kehinde
There is some clinical evidence to suggest that fractures of long bones heal more rapidly in patients with severe head injury or acute traumatic spinal cord injury. The mechanism underlying this orthopedic phenomenon is not well understood. The aim of the current study was to investigate the bone healing of diaphyseal femoral fractures in patients with concomitant head or spinal cord injuries and to elucidate mechanism of a possible accelerated osteogenesis. The study recruited 52 group (A) patients with head injury, 50 group (B) patients with head injury and 58 femoral shaft fractures, 20 group (C) patients with spinal cord injuries, 21 group (D) patients with spinal cord injuries and 22 femoral shaft fractures, 60 group (E) patients with 69 femoral shaft fractures only, and 50 group (F) healthy subjects. All the femoral fractures in groups (B), (D), and (E) were treated by closed reduction and internal fixation by intramedullary nail. The fracture healing indicators were compared between patients of different groups and patients' blood samples were used to count circulating mesenchymal stem cells (MSCs) and monocytes using the flow cytometry. Results were analysed with statistical package of social sciences SPSS. Mean scores between two groups of patients were compared using chi square and the Student t-test. The study showed that femoral diaphyseal fractures in patients with head or spinal cord injury heal more expectedly, faster and with more callus formation and patients' blood samples showed statistically significant increase in circulating MSCs count and blood monocytes count in groups (A) to (D). The study revealed acceleration of femoral fractures in patients with concomitant head or spinal cord injuries and also demonstrated mobilization of distant bone marrow MSCs, homing early to fracture site and the role of monocytes in providing mediators to accelerate healing.
{"title":"Estimation of mesenchymal stem cells (mscs) and monocytes in peripheral blood of acute traumatic head and spinal cord injuries in patients with concomitant diaphyseal femoral fractures: a clinical prospective controlled cohort study","authors":"F. Khallaf, E. Kehinde","doi":"10.15406/JSRT.2016.01.00043","DOIUrl":"https://doi.org/10.15406/JSRT.2016.01.00043","url":null,"abstract":"There is some clinical evidence to suggest that fractures of long bones heal more rapidly in patients with severe head injury or acute traumatic spinal cord injury. The mechanism underlying this orthopedic phenomenon is not well understood. The aim of the current study was to investigate the bone healing of diaphyseal femoral fractures in patients with concomitant head or spinal cord injuries and to elucidate mechanism of a possible accelerated osteogenesis. The study recruited 52 group (A) patients with head injury, 50 group (B) patients with head injury and 58 femoral shaft fractures, 20 group (C) patients with spinal cord injuries, 21 group (D) patients with spinal cord injuries and 22 femoral shaft fractures, 60 group (E) patients with 69 femoral shaft fractures only, and 50 group (F) healthy subjects. All the femoral fractures in groups (B), (D), and (E) were treated by closed reduction and internal fixation by intramedullary nail. The fracture healing indicators were compared between patients of different groups and patients' blood samples were used to count circulating mesenchymal stem cells (MSCs) and monocytes using the flow cytometry. Results were analysed with statistical package of social sciences SPSS. Mean scores between two groups of patients were compared using chi square and the Student t-test. The study showed that femoral diaphyseal fractures in patients with head or spinal cord injury heal more expectedly, faster and with more callus formation and patients' blood samples showed statistically significant increase in circulating MSCs count and blood monocytes count in groups (A) to (D). The study revealed acceleration of femoral fractures in patients with concomitant head or spinal cord injuries and also demonstrated mobilization of distant bone marrow MSCs, homing early to fracture site and the role of monocytes in providing mediators to accelerate healing.","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"4 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79012848","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-11-29DOI: 10.15406/jsrt.2016.01.00042
Seyed Meysam Abtahi Froushani
Regenerative or reparative medicine isa new term which emerged in medicine to refer to renewing of damaged organs by stem cells. Mesenchymal stem cellsare multipotent progenitor cells that have the capacity to differentiate into all lineages of mesodermal origin. Mesenchymal stem cells (MSCs) have a good potential for inducing anti-inflammatory responses in inflamed tissues, hence they are used as therapy in auto-inflammatory disease. Due to their differentiation, proliferation and self- renewing characteristics, have received attentions with regard to their potential use as therapeutic agent. This promising area of science is leading scientists to investigate the possibility of cell-based therapies to treat different kinds of diseases, e.g., GVHD, Multiple sclerosis, diabetes and etc. There are two population of cells in the bone marrow that can be differentiated to MSCs in vitro.Mesenchymal stem cells derived from the non-adherent cell population of human bone marrow cell cultures had similar cell proliferation rates in vitrowhen compared with the MSCs derived from the primary adherent cell population. While there are different methods for isolation and preparation of MSCs from different tissues, but none of them are completely standard. All these methods need additional manipulations of cells, which may affect their differentiation potentials as well as increasing the risk of contamination of cultures. Therefore, there is a need to compare these populations from different aspects. We hypothesize that non adherent MSCs population are more applicable than adherent one, therefore, culturing and manipulating of non-adherent MSCs population is easier than adherent MSCs. In addition, non-adherent MSCs culturing is a cost-effective way as well as increasing the number of cells and shortening the time of the cell culture.
{"title":"Adherent or non-adherent mesenchymal stem cell; which one is more applicable for clinical study?","authors":"Seyed Meysam Abtahi Froushani","doi":"10.15406/jsrt.2016.01.00042","DOIUrl":"https://doi.org/10.15406/jsrt.2016.01.00042","url":null,"abstract":"Regenerative or reparative medicine isa new term which emerged in medicine to refer to renewing of damaged organs by stem cells. Mesenchymal stem cellsare multipotent progenitor cells that have the capacity to differentiate into all lineages of mesodermal origin. Mesenchymal stem cells (MSCs) have a good potential for inducing anti-inflammatory responses in inflamed tissues, hence they are used as therapy in auto-inflammatory disease. Due to their differentiation, proliferation and self- renewing characteristics, have received attentions with regard to their potential use as therapeutic agent. This promising area of science is leading scientists to investigate the possibility of cell-based therapies to treat different kinds of diseases, e.g., GVHD, Multiple sclerosis, diabetes and etc. There are two population of cells in the bone marrow that can be differentiated to MSCs in vitro.Mesenchymal stem cells derived from the non-adherent cell population of human bone marrow cell cultures had similar cell proliferation rates in vitrowhen compared with the MSCs derived from the primary adherent cell population. While there are different methods for isolation and preparation of MSCs from different tissues, but none of them are completely standard. All these methods need additional manipulations of cells, which may affect their differentiation potentials as well as increasing the risk of contamination of cultures. Therefore, there is a need to compare these populations from different aspects. We hypothesize that non adherent MSCs population are more applicable than adherent one, therefore, culturing and manipulating of non-adherent MSCs population is easier than adherent MSCs. In addition, non-adherent MSCs culturing is a cost-effective way as well as increasing the number of cells and shortening the time of the cell culture.","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"80 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83867756","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-11-23DOI: 10.15406/JSRT.2016.01.00041
Hassan Dana, Vahid Marmari, Ghanbar Mahmoodi, H. Mahmoodzadeh, M. Ebrahimi, Narges Mehm, Oost
CD166 (Activated leukocyte cell adhesion molecule (ALCAM)) is a member of the immunoglobulin superfamily, which expressed by various cells in several tissues including colorectal cancer (CRC). Many studies have reported the prognostic predictive value of CD166 as a cancer stem cell marker in CRC. CD166 gained increasing attention regarding tumor progression and metastatic spread in CRC. CD166 potentially represents either diagnostic or therapeutic capacities for CRC. This Mini review aimed to clarify the role and significance of CD166 expression in CRC.
{"title":"CD166 as a stem cell marker? a potential target for therapy colorectal cancer?","authors":"Hassan Dana, Vahid Marmari, Ghanbar Mahmoodi, H. Mahmoodzadeh, M. Ebrahimi, Narges Mehm, Oost","doi":"10.15406/JSRT.2016.01.00041","DOIUrl":"https://doi.org/10.15406/JSRT.2016.01.00041","url":null,"abstract":"CD166 (Activated leukocyte cell adhesion molecule (ALCAM)) is a member of the immunoglobulin superfamily, which expressed by various cells in several tissues including colorectal cancer (CRC). Many studies have reported the prognostic predictive value of CD166 as a cancer stem cell marker in CRC. CD166 gained increasing attention regarding tumor progression and metastatic spread in CRC. CD166 potentially represents either diagnostic or therapeutic capacities for CRC. This Mini review aimed to clarify the role and significance of CD166 expression in CRC.","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"56 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91440811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-11-21DOI: 10.15406/JSRT.2016.01.00039
P. Deeksha, Ey, Bhanu Pratap Singh Dhakar, Vibhor Ahluwalia
This article focuses the pros and cons of various UCB banking models. We also discuss here the recommendation by various medical societies in this regard as well as the future direction in the subject of UCB banking. On the practical grounds education of general population, obstetrical care providers and paediatricians on the current banking strategies and potential uses of UCB is essential. While providing this education it has to be kept in mind the limited number of public UCB banks in many countries across the world; declaring an urgent need to simultaneously have government policies to make such banking services available to the potential donors.
{"title":"Umbilical Cord Blood (UCB) Banking: Which One to Choose?","authors":"P. Deeksha, Ey, Bhanu Pratap Singh Dhakar, Vibhor Ahluwalia","doi":"10.15406/JSRT.2016.01.00039","DOIUrl":"https://doi.org/10.15406/JSRT.2016.01.00039","url":null,"abstract":"This article focuses the pros and cons of various UCB banking models. We also discuss here the recommendation by various medical societies in this regard as well as the future direction in the subject of UCB banking. On the practical grounds education of general population, obstetrical care providers and paediatricians on the current banking strategies and potential uses of UCB is essential. While providing this education it has to be kept in mind the limited number of public UCB banks in many countries across the world; declaring an urgent need to simultaneously have government policies to make such banking services available to the potential donors.","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75111302","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-11-17DOI: 10.15406/jsrt.2016.01.00038
B. Mehling, M. Manvelyan, G. Benesh, Dongcheng Wu
Medium where the stem cells are cultured is called conditioned medium (CM). In CM, Mesenchymal Stem Cells (MSCs) secrete different growth factors and cytokines with anti-inflammatory and anti-fibrotic effects. Numerous questions need to be answered before MSCs-derived CM can be used for the therapy of various diseases/conditions. The purpose of the present study is to analyze Human Umbilical Cord MSCs-derived CM for the presence of anti-inflammatory and regeneration-promoting cytokines. In our study of CM, immunoassay tests showed that all analyzed chemokines gave signals in the assays in positive control parallel wells. Thus all chemokines are functional and accurate to the indicated levels. Characterization of growth factors and cytokines in the MSCs-derived CM is crucial for further translation of CM in therapy of various diseases/conditions.
{"title":"Characterization of Human Umbilical Cord Mesenchymal Stem Cells-Derived Conditioned Medium","authors":"B. Mehling, M. Manvelyan, G. Benesh, Dongcheng Wu","doi":"10.15406/jsrt.2016.01.00038","DOIUrl":"https://doi.org/10.15406/jsrt.2016.01.00038","url":null,"abstract":"Medium where the stem cells are cultured is called conditioned medium (CM). In CM, Mesenchymal Stem Cells (MSCs) secrete different growth factors and cytokines with anti-inflammatory and anti-fibrotic effects. Numerous questions need to be answered before MSCs-derived CM can be used for the therapy of various diseases/conditions. The purpose of the present study is to analyze Human Umbilical Cord MSCs-derived CM for the presence of anti-inflammatory and regeneration-promoting cytokines. In our study of CM, immunoassay tests showed that all analyzed chemokines gave signals in the assays in positive control parallel wells. Thus all chemokines are functional and accurate to the indicated levels. Characterization of growth factors and cytokines in the MSCs-derived CM is crucial for further translation of CM in therapy of various diseases/conditions.","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88103543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-11-17DOI: 10.15406/JSRT.2016.01.00037
Carina Adamzyk, N. Labude, R. Schneider, J. Jaekel, Hoffmann, J. Fischer, M. Hoss, R. Knuechel, W. JahnenDechent, S. Neuss
Human adult mesenchymal stem cells (hMSC) are particularly suitable cells for autologous tissue engineering and cell-based therapies. They can be isolated from various tissues, such as bone marrow, adipose tissue, dental pulp or umbilical cords. Due to their primitive developmental stage, umbilical cord-derived hMSC are assumed to have a higher proliferation and differentiation capacity than hMSC from adult tissues. We isolated hMSC from bone marrow (BM) and umbilical cords (UC) and compared the cells regarding their surface epitopes, proliferation and differentiation capacity. Flow cytometry of specific surface epitopes showed that both BM-MSC and UC-MSC display the characteristic MSC phenotype. Cells from both sources were readily differentiated into adipocytes, osteoblasts and chondrocytes according to standard protocols. Interestingly, only UC-MSC spontaneously formed three-dimensional aggregates when cultured under post-confluent conditions. The cells of these aggregates were viable and spontaneously differentiated into several specialized cell types akin to the well-known differentiation of embryoid bodies. Besides, UC-MSC expressed the pluripotency-associated gene NANOG as well as genes characteristic for the mesodermal and ectodermal fate. Thus, UC-MSC resemble BM-MSC but additionally show a spontaneous embryoid body-like aggregation and differentiation in vitro. These results indicate that UC-MSC are less restricted than BM-MSC and may thus extend the limits of BM-MSC based therapies.
{"title":"Human Umbilical Cord-Derived Mesenchymal Stem Cells Spontaneously Form 3D Aggregates and Differentiate in an Embryoid Body-Like Manner","authors":"Carina Adamzyk, N. Labude, R. Schneider, J. Jaekel, Hoffmann, J. Fischer, M. Hoss, R. Knuechel, W. JahnenDechent, S. Neuss","doi":"10.15406/JSRT.2016.01.00037","DOIUrl":"https://doi.org/10.15406/JSRT.2016.01.00037","url":null,"abstract":"Human adult mesenchymal stem cells (hMSC) are particularly suitable cells for autologous tissue engineering and cell-based therapies. They can be isolated from various tissues, such as bone marrow, adipose tissue, dental pulp or umbilical cords. Due to their primitive developmental stage, umbilical cord-derived hMSC are assumed to have a higher proliferation and differentiation capacity than hMSC from adult tissues. We isolated hMSC from bone marrow (BM) and umbilical cords (UC) and compared the cells regarding their surface epitopes, proliferation and differentiation capacity. \u0000 \u0000 Flow cytometry of specific surface epitopes showed that both BM-MSC and UC-MSC display the characteristic MSC phenotype. Cells from both sources were readily differentiated into adipocytes, osteoblasts and chondrocytes according to standard protocols. Interestingly, only UC-MSC spontaneously formed three-dimensional aggregates when cultured under post-confluent conditions. The cells of these aggregates were viable and spontaneously differentiated into several specialized cell types akin to the well-known differentiation of embryoid bodies. Besides, UC-MSC expressed the pluripotency-associated gene NANOG as well as genes characteristic for the mesodermal and ectodermal fate. Thus, UC-MSC resemble BM-MSC but additionally show a spontaneous embryoid body-like aggregation and differentiation in vitro. These results indicate that UC-MSC are less restricted than BM-MSC and may thus extend the limits of BM-MSC based therapies.","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"39 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84270992","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2016-11-10DOI: 10.15406/JSRT.2016.01.00036
S. K. Varma
Cancer is one of the biggest challenges of modern medicine. Though there are different therapies in general like chemotherapy, radiation and surgery. Further, cancer is highly heterogeneous. Two persons, who suffer from the same type of cancer, can show totally different variations and ways of progress. That makes it hard to treat the cancer and just for that reason oncology is one field where personalized medicine is advancing extremely fast. The more that treatments can be tailored to the single patient, the higher are the chances of treating that patient’s cancer effectively. Personalization creating a special tailored therapy for everybody wasn’t possible before. The goal is to allow patients a pain free time at least for a while, to improve the quality of life and to dam up the growth of the tumor. To reach that goal, now researchers concentrate on the power of the human immune system and its improvement to treat cancers with immunotherapy. Dendritic cells (DCs) play a central role in the initiation and regulation of innate and adaptive immune responses and have increasingly been applied as vaccines for cancer patients. In vitro generation of dendritic cells from monocytes and antigen loading into immature dendritic cells to proper maturation, with the aim of imprinting different DC functions that are essential for their subsequent induction of a T cell-mediated immune response against tumor.
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Pub Date : 2016-11-04DOI: 10.15406/JSRT.2016.01.00035
Guang-Jer Wu
METCAM/MUC18, an integral membrane cell adhesion molecule (CAM) in the Ig-like gene super family, is not expressed in most normal prostate gland, or in all BPH, but overly expressed in most malignant prostate cancer. Its overexpression also correlates with the malignant progression of mouse prostatic adenocarcinoma in a TRAMP model, suggesting that it may be a diagnostic marker for malignant prostate cancer. To demonstrate this, three immunological methods are developed to show promises to use METCAM/MUC18 as a diagnostic marker for the presence of clinical prostate cancer. Enforced expression of METCAM/ MUC18 in a human prostate cancer cell line, LNCaP, promotes tumorigenesis and initiates metastasis to multiple organs in a male nude mouse model, suggesting that it can promote the malignant progression of prostate cancer. ShRNAs in a lentivirus vector could decrease the tumorigenesis of another human prostate cancer cell line, DU145, in a male athymic nude mouse model. Taken together, METCAM/MUC18 is a new diagnostic marker of and a driver for promoting the malignant progression of and its specific siRNAs, oligo-peptides, and antibodies may be used for therapeutic treatments of clinical prostate cancer.
{"title":"Human METCAM/MUC18 is a New Diagnostic Marker of and a Driver for Promoting and its Specific SiRNAs, Derived Oligopeptides and Antibodies be used for Decreasing the Malignant Progression of Prostate Cancer","authors":"Guang-Jer Wu","doi":"10.15406/JSRT.2016.01.00035","DOIUrl":"https://doi.org/10.15406/JSRT.2016.01.00035","url":null,"abstract":"METCAM/MUC18, an integral membrane cell adhesion molecule (CAM) in the Ig-like gene super family, is not expressed in most normal prostate gland, or in all BPH, but overly expressed in most malignant prostate cancer. Its overexpression also correlates with the malignant progression of mouse prostatic adenocarcinoma in a TRAMP model, suggesting that it may be a diagnostic marker for malignant prostate cancer. To demonstrate this, three immunological methods are developed to show promises to use METCAM/MUC18 as a diagnostic marker for the presence of clinical prostate cancer. Enforced expression of METCAM/ MUC18 in a human prostate cancer cell line, LNCaP, promotes tumorigenesis and initiates metastasis to multiple organs in a male nude mouse model, suggesting that it can promote the malignant progression of prostate cancer. ShRNAs in a lentivirus vector could decrease the tumorigenesis of another human prostate cancer cell line, DU145, in a male athymic nude mouse model. Taken together, METCAM/MUC18 is a new diagnostic marker of and a driver for promoting the malignant progression of and its specific siRNAs, oligo-peptides, and antibodies may be used for therapeutic treatments of clinical prostate cancer.","PeriodicalId":91560,"journal":{"name":"Journal of stem cell research & therapeutics","volume":"28 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76625377","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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