Large‐scale manufacturing of safe and efficient retrovirus packaging lines for use in immunotherapy protocols

Farson, Deborah; McGuinness, Ryan; Dull, Tom; Limoli, Kay; Lazar, Richard; Jalali, Sayeh; Reddy, Sridhar; Pennathur-Das, Rukmini; Broad, David; Finer, Mitchell

doi:10.1002/(sici)1521-2254(199905/06)1:3<195::aid-jgm31>3.0.co;2-#

Cited by 25 publications

(10 citation statements)

References 49 publications

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“…Tissue culture medium from transfected 293-10A1 cells [42] was filtered 48 h after transfection, and the viral supernatant was used to infect cultures of the subconfluent ECC-1 cells after the addition of 4 lg/ml polybrene. Isolated clones were obtained using antibiotic selection (puromycin) and further expanded to confluence to obtain stably transfected cells.…”

Section: Knockdown Of Muc16 and Muc1 In Ecc-1 Cellsmentioning

confidence: 99%

MUC16 Is Lost from the Uterodome (Pinopode) Surface of the Receptive Human Endometrium: In Vitro Evidence That MUC16 Is a Barrier to Trophoblast Adherence1

Gipson

Blalock

Tisdale

et al. 2008

Biology of Reproduction

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In order for the preimplantation embryo to implant into the uterus, the trophoblast cells must initially adhere to the uterine epithelial surface. In preparation, the luminal secretory cells of the epithelium lose their nonadhesive character and their surface microvilli and bulge into the lumen, forming uterodomes (pinopodes; uterodome is used instead of pinopode, since in humans the surface membrane exocytoses rather than endocytoses (Murphy, Hum Reprod 2000; 15:2451-2454). Previous research has led to the hypothesis that loss of the nonadhesive membrane-spanning mucin MUC1 from the uterodome surface allows trophoblast adherence. Immunofluorescence microscopic assay of luminal epithelia on human uterine biopsies taken from LH+0 to LH+13 show that another membrane-spanning mucin, MUC16, was lost from uterodome surfaces in all samples taken during the receptive phase, LH+6 to LH+8 (n = 12), and that MUC1 was present on uterodomes in 4 of 12 samples and on all ciliated cells of the epithelium in the receptive phase. Short interfering RNA (siRNA) knockdown of MUC16 in a uterine epithelial cell line ECC-1 that, like uterine epithelium, expresses MUC16 and MUC1 allowed increased adherence of cells of a trophoblast cell line. In parallel experiments, siRNA knockdown of MUC1 did not affect trophoblast cell adherence. These data indicate that MUC16 is a membrane component of the nonreceptive luminal uterine surface, which prevents cell adhesion, and that its removal during uterodome formation facilitates adhesion of the trophoblast.

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Section: Knockdown Of Muc16 and Muc1 In Ecc-1 Cellsmentioning

confidence: 99%

MUC16 Is Lost from the Uterodome (Pinopode) Surface of the Receptive Human Endometrium: In Vitro Evidence That MUC16 Is a Barrier to Trophoblast Adherence1

Gipson

Blalock

Tisdale

et al. 2008

Biology of Reproduction

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“…Although many different culture systems have been tested and described (for details, the reader is referred to the review by Merten [31]) only very few systems have really been used for the production of clinical grade MLV vectors. Smaller preclinical and clinical grade quantities were generally produced with roller bottles [28,30] or CFs [29,32,34,51], whereas large vector preparations have been produced using fixed bed bioreactors (CellCube) [30,34,[90][91] as well as Wave bioreactor [92] Although roller bottles and CFs could be used for more than one week (see above) all large-scale productions were limited to the harvests on 3-4 consecutive days (Table 1). For both production systems, depending on the number of culture units the overall harvest volume could be adapted to the needs.…”

Section: Production Systems For the Large Scale Manufacturing Of MLV mentioning

confidence: 99%

Manufacturing of viral vectors for gene therapy: part I. Upstream processing

Merten¹,

Schweizer²,

Chahal³

et al. 2014

Pharmaceutical Bioprocessing

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“…For this purpose, mobilized peripheral blood (MPB)-derived CD34 1 cells were cultured in a transwell system that contained in the lower chamber either the murine cell line NIH 3T3 (Todaro and Green, 1963) (basal cell for GP1Am12; Markowitz et al, 1988), the murine BM stromal cell line MS-5 (Itoh et al, 1989), the human cell line HEK 293 (Graham et al, 1977) (basal cell for ProPak and Allikat [Farson et al, 1999]), the human cell line ECV 304/T24 (Takahashi et al, 1990), or primary human BM stroma. After 7 and 14 days of culture in medium without any exogenous cytokines, maintenance of CD34 1 cells was evaluated by flow cytometric analysis counts.…”

Section: Ecv 304/t24 Maintains Human Cd34 1 Cells At a Level Equivalementioning

confidence: 99%

“…The use of human packaging cells has revolved around two cell lines, the transformed embryonic kidney epithelial cell line HEK 293 Farson et al, 1999) and the renal fibrosarcoma HT 1080 (Cosset et al, 1995). Both have been reported to give rise to efficient levels of hematopoietic cell transduction (Onodera et al, 1998;Murray et al, 1999), but neither has been reported to have hematopoietic effector function or to constitutively secrete known hematopoietically acting cytokines or growth factors.…”

Section: Introductionmentioning

confidence: 99%

A Novel Human Packaging Cell Line with Hematopoietic Supportive Capacity Increases Gene Transfer into Early Hematopoietic Progenitors

et al. 2001

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The hematopoietic stem/progenitor cell (HSPC) represents the ideal target for gene therapy of disorders of the hematopoietic system, but still faces problems related to ex vivo manipulation and gene transfer efficiency. We demonstrate that soluble factors from the human endothelial-like cell line ECV 304/T24 support the growth of human CD34(+) progenitor cells as primary human bone marrow stroma and increase the rate of gene transfer into progenitor cells up to 5-fold. ECV 304/T24 was used to generate split-function amphotropic packaging cell lines (named APEX) with the purpose of combining, in the same cells, hematopoietic support and gene transfer vehicle functions. The APEX cell lines were negative for the presence of replication-competent retroviruses and produced complement-resistant vector particles. When mobilized peripheral blood or umbilical cord blood CD34(+) cells were exposed once to APEX supernatants, the level of gene transfer was equivalent to that observed with GP + Am12, in spite of the lower titer of the APEX producers. More importantly, APEX supernatants gave rise reproducibly to a 2-fold increase in transduction of early progenitors (long-term culture-initiating cells), reaching on average 50% gene transfer. This novel packaging cell represents a significant advance in HSPC genetic modification technology, combining both a beneficial hematopoietic supportive effect and the gene transfer vector function in a human-based system.

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Large‐scale manufacturing of safe and efficient retrovirus packaging lines for use in immunotherapy protocols

Cited by 25 publications

References 49 publications

MUC16 Is Lost from the Uterodome (Pinopode) Surface of the Receptive Human Endometrium: In Vitro Evidence That MUC16 Is a Barrier to Trophoblast Adherence1

MUC16 Is Lost from the Uterodome (Pinopode) Surface of the Receptive Human Endometrium: In Vitro Evidence That MUC16 Is a Barrier to Trophoblast Adherence1

Manufacturing of viral vectors for gene therapy: part I. Upstream processing

A Novel Human Packaging Cell Line with Hematopoietic Supportive Capacity Increases Gene Transfer into Early Hematopoietic Progenitors

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