Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/W mice

Dick, John E.; Magli, Maria Cristina; Huszar, Dennis; Phillips, Robert A.; Bernstein, Alan

doi:10.1016/s0092-8674(85)80102-1

Cited by 625 publications

(277 citation statements)

References 24 publications

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“…22,31 After red blood cell ( RBC ) depletion with PharM Lyse 2 ( PharMingen, Heidelberg, Germany ), according to the manufacturer's instructions, cells were prestimulated for 48 hours in -modified Eagle's medium (Sigma ) containing 20% FCS, P/ S, 100 U / mL recombinant human (rh ) IL -6 (Novartis, Basel, Switzerland ), and 50 ng/ mL recombinant murine ( rm ) SCF ( PeproTech, Rocky Hill, NJ, USA ). Transduction of prestimulated bone marrow was carried out in 10 -cm nontissue culture Petri dishes (Becton Dickinson, Heidelberg, Germany ) coated with 4 g/cm 2 retronectin (BioWhittaker, Taufkirchen, Germany ), as described previously.…”

Section: Bone Marrow Preparation Transduction and Transplantationmentioning

confidence: 99%

Hematoprotection and enrichment of transduced cells in vivo after gene transfer of MGMTP140K into hematopoietic stem cells

et al. 2002

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The overexpression of mutant forms of O 6 -methylguanine -DNA -methyltransferase ( MGMT ), resistant to the MGMT inhibitor O 6 -benzylguanine ( BG ), protects hematopoietic cells from the toxicity of combined BG plus O 6 -alkylating agent chemotherapy. To evaluate the feasibility of this approach for clinically relevant O 6 -alkylating agents, combined therapy with BG and two chloroethylnitrosourea -type drugs, ACNU or BCNU, or the triazene derivative temozolomide ( TMZ ) was investigated in a murine bone marrow transplant model allowing transgenic expression of the highly BG -resistant MGMT P140K mutant. Whereas 20 / 20 control animals transplanted with nontransduced cells died of progressive myelosuppression during therapy, nearly all animals transplanted with MGMT P140K -transduced cells survived treatment with BG / ACNU ( 12 / 15 ), BG / TMZ ( 10 / 10 ), or BG / BCNU (5/ 5 ). In surviving animals, hematological parameters improved during chemotherapy and pretreatment levels were reestablished during or shortly after therapy. All animals showed enrichment of transgenic granulocytes ( range: 15 -to 101 -fold ) and lymphocytes ( range: 16 -to 55 -fold ) in peripheral blood, bone marrow, and spleen. No significant differences were observed between individual treatment groups. Serial transplants demonstrated protection in secondary recipients and confirmed the transduction of transplantable stem cells. Thus, these data demonstrate efficient protection from hematotoxicity and substantial enrichment of transgenic cells following MGMT P140K gene transfer and treatment with different O 6 -alkylating drugs.

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Section: Bone Marrow Preparation Transduction and Transplantationmentioning

confidence: 99%

Hematoprotection and enrichment of transduced cells in vivo after gene transfer of MGMTP140K into hematopoietic stem cells

et al. 2002

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“…[1][2][3] The mechanisms that regulate the early stages of murine B-lymphopoiesis from primitive hematopoietic progenitors have been extensively studied. Cumano et al 4 developed a stroma celldependent clonal culture assay in which bipotential progenitors from murine day-12 fetal liver differentiated along Blymphoid and macrophage lineages in the presence of stromal cells and interleukin (IL)-7.…”

Section: Introductionmentioning

confidence: 99%

Culture system for extensive production of CD19+IgM+ cells by human cord blood CD34+ progenitors

et al. 1998

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“…[22][23][24][25][26][27][28][29][30][31][32] Dominant selectable markers include naturally occurring human drug resistance genes such as MDR1, DHFR L22Y or DNA alkyltransferases. 22,23,25,26,[31][32][33][34] This approach requires the use of a vector encoding both the gene of interest and the dominant selectable marker.…”

Section: Marked Cells In the Presence Of The Antifolate Drug Trimetrementioning

confidence: 99%

Pre-clinical evaluation of an in vitro selection protocol for the enrichment of transduced CD34+ cell-derived human dendritic cells

et al. 2001

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The efficient genetic modification of CD34 + cell-derived dendritic cells (DC) will provide a significant advancement towards the development of immunotherapy protocols for cancer, autoimmune disorders and infectious diseases. Recent reports have described the transduction of CD34 + cells via retrovirus-and lentivirus-based gene transfer vectors and subsequent differentiation into functional DC. Since there is significant apprehension regarding the clinical uses of HIV-based vectors, in this report, we compare a murine leukemia virus (MLV)-and a human immunodeficiency virus (HIV)-based bicistronic vector for gene transfer into human

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Introduction of a selectable gene into primitive stem cells capable of long-term reconstitution of the hemopoietic system of W/W mice

Cited by 625 publications

References 24 publications

Hematoprotection and enrichment of transduced cells in vivo after gene transfer of MGMTP140K into hematopoietic stem cells

Hematoprotection and enrichment of transduced cells in vivo after gene transfer of MGMTP140K into hematopoietic stem cells

Culture system for extensive production of CD19+IgM+ cells by human cord blood CD34+ progenitors

Pre-clinical evaluation of an in vitro selection protocol for the enrichment of transduced CD34+ cell-derived human dendritic cells

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