High efficiency retroviral mediated gene transduction into single isolated immature and replatable CD34(3+) hematopoietic stem/progenitor cells from human umbilical cord blood.

Lü, Li; Xiao, Mang; Clapp, D. Wade; Li, Zhihua; Broxmeyer, Hal E.

doi:10.1084/jem.178.6.2089

Cited by 106 publications

(54 citation statements)

References 34 publications

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“…15 Finally, gene transduction protocols have been more successful with CB cells than with BM or mPB cells. 9 In the current study, CFU-GM expansion (126-fold) obtained with CB cells at an inoculum density of 3 ϫ 10 5 per cm 2 was significantly greater than that obtained with adult BM cells (13-fold) under identical culture conditions. 27 These different lines of evidence indicate that CB cell populations are significantly more primitive and have more in vitro and in vivo proliferative potential than BM or mPB cells.…”

Section: Discussioncontrasting

confidence: 44%

“…In fact, many studies have suggested that CB contains a more primitive cell population, having more in vitro and in vivo proliferative potential than BM or mPB cells, [8][9][10][11][12][13][14][15] making them a prime target for ex vivo expansion therapy or gene therapy procedures. Most of the recent literature on CB cell expansion has focused on the culture of CD34-enriched cells or subsets thereof.…”

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confidence: 99%

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Clinical-scale human umbilical cord blood cell expansion in a novel automated perfusion culture system

Koller

Manchel

Maher

et al. 1998

Bone Marrow Transplant

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Summary:Use of umbilical cord blood (CB) for stem cell transplantation has a number of advantages, but a major disadvantage is the relatively low cell number available. Ex vivo cell expansion has been proposed to overcome this limitation, and this study therefore evaluated the use of perfusion culture systems for CB cell expansion. CB was cryopreserved using standard methods and the thawed unpurified cells were used to initiate small-scale cultures supplemented with PIXY321, flt-3 ligand, and erythropoietin in serum-containing medium. Twelve days of culture resulted in the optimal output from most CB samples. Frequent medium exchange led to significant increases in cell (93%), CFU-GM (82%) and LTC-IC (350%) output as compared with unfed cultures. As the inoculum density was increased from 7.5 ؋ 10 4 per cm 2 to 6.0 ؋ 10 5 per cm 2 , the output of cells, CFU-GM, and LTC-IC increased. Cell and CFU-GM output reached a plateau at 6.0 ؋ 10 5 per cm 2 , whereas LTC-IC output continued to increase up to 1.2 ؋ 10 6 per cm 2 . Because the increase in culture output did not increase linearly with increasing inoculum density, expansion ratios were greatest at 1.5 ؋ 10 5 per cm 2 for cells (6.4-fold) and CFU-GM (192-fold). Despite the lack of adherent stroma, CB cultures expressed mRNA for many growth factors (G-CSF, GM-CSF, IL-1, IL-6, LIF, KL, FL, Tpo, TGF-␤, TNF-␣, and MIP-1␣) that were also found in bone marrow (BM) cultures, with the exception of IL-11 (found only in BM) and IL-3 (found in neither). Culture output was remarkably consistent from 10 CB samples (coefficient of variation 0.3) indicating that the procedure is robust and reproducible. Two commercial serum-free media were evaluated and found to support only approximately 25% of the culture output as compared with serum-containing medium. Implementation of optimal conditions in the clinical scale, automated cell production system (CPS) showed that the process scaled-up well, generating 1.7 ؋ 10 7 CFU-GM (298-fold expansion) from 1.2 ؋ 10 8 thawed viable nucleated CB cells (n = 3). The ability to generate Ͼ Ͼ Ͼ10 7 CFU-GM from Ͻ15 ml of CB within this closed, automated system without the need for extensive cell manipulations

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Section: Discussioncontrasting

confidence: 44%

mentioning

confidence: 99%

Clinical-scale human umbilical cord blood cell expansion in a novel automated perfusion culture system

Koller

Manchel

Maher

et al. 1998

Bone Marrow Transplant

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“…This combination also gave improved transduction frequencies with a retroviral vector encoding the Fanconi's anaemia group C protein 24 and using a neoR reporter. 25 Other successful combinations have included SCF, IL-3, GM-CSF, Epo 26 and SCF, Flt3-ligand, GM-CSF, TNF-␣, TGF-␤. 27 The system we have described in this report lends itself well to the transduction of populations of cells like these that are predominantly quiescent but without recourse to the use of exogenously added cytokines.…”

Section: Discussionmentioning

confidence: 99%

Retroviral transduction of quiescent haematopoietic cells using a packaging cell line expressing the membrane-bound form of stem cell factor

1999

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Gene therapy vectors based on murine retroviruses are strated that these cells, though quiescent, can still be sucunable to transduce non-dividing cells. This has proven a cessfully transduced. This approach was extended to tarparticular problem in the haematopoietic system where the geting of umbilical cord blood CD34 + cells, a predominantly target cells of choice, the pluripotent stem cells are quiescquiescent population that normally require the addition of ent. In an attempt to circumvent this difficulty we have concytokines for efficient transduction. Using the SCFstructed a retroviral producer line that expresses the memexpressing producer line in the absence of exogenously brane bound form of human recombinant stem cell factor added cytokines, we observed a marked stimulation in (SCF) on its cell surface. This should enable the retroviral transduction efficiency over that achieved using the parent producers to deliver a growth signal to the target cells simproducer line alone. Colonies derived from these cells arisultaneous with their exposure to retrovirus. We tested the ing in semi-solid media were also shown to be positive for ability of these modified producers to transduce a growth expression of a retrovirally encoded reporter gene. factor-starved, SCF-dependent cell line (TF-1) and demon-

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“…In vitro studies on PUCB CD34 + cells have indicated that retrovirus-transduced genes can be introduced into 12-95% of progenitor cells. 9,[11][12][13] In vivo animal studies using SCID mouse models have demonstrated that CD34 + cells derived from PUCB could be successfully engrafted. 14 These promising data have recently led to the initiation of a human gene therapy trial using PUCB-derived CD34 + cells for adenosine deaminase deficiency.…”

Section: Discussionmentioning

confidence: 99%

Gene therapy targeting cord blood-derived CD34+ cells from HIV-exposed infants: preclinical studies

Gervaix

Kang

et al. 1998

Gene Ther

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Hematopoietic CD34+ cells from placental and umbilical cord blood (PUCB) can be valuable vehicles for gene therapy of immunodeficiencies and genetic disorders. We have conducted preclinical studies towards the treatment of HIV-1-infected infants with genetically 'immunized' CD34+ cells derived from PUCB using anti-HIV-1 hairpin ribozyme genes. PUCB was collected from 10 newborns of HIV-1-positive mothers. CD34+ cells were enriched with a modified procedure using Dynal immunomagnetic beads and chymopapain, stimulated with stem cell factor (SCF), IL-3 and IL-6, and transduced using cell-free recombinant retroviral vector (MJT) expressing a ribozyme against the U5 region of HIV-1. No significant differences were observed in the growth pattern of CD34+ cells from normal infants, HIV-1 exposed infants or infants confirmed to be infected by HIV-1. The transduction efficiency of the CD34+ cells from all the infants was also comparable. MJT-transduced CD34+ cells from an HIV-1-infected infant were maintained in a liquid culture system for 4 weeks, and the progeny macrophage cells were challenged with the monocyte-tropic laboratory strain, HIV-Bal, or the HIV-1 isolate from the infant's mother. Significant inhibition of virus replication was observed in ribozyme-transduced cells. Thus, we have demonstrated efficient and stable gene transfer into progenitor cells from the cord blood of HIV-1-exposed or -infected infants and shown that protection from HIV-1 infection was conferred to the progeny cells produced by CD34+ cells transduced with the anti-HIV ribozyme gene construct

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High efficiency retroviral mediated gene transduction into single isolated immature and replatable CD34(3+) hematopoietic stem/progenitor cells from human umbilical cord blood.

Cited by 106 publications

References 34 publications

Clinical-scale human umbilical cord blood cell expansion in a novel automated perfusion culture system

Clinical-scale human umbilical cord blood cell expansion in a novel automated perfusion culture system

Retroviral transduction of quiescent haematopoietic cells using a packaging cell line expressing the membrane-bound form of stem cell factor

Gene therapy targeting cord blood-derived CD34+ cells from HIV-exposed infants: preclinical studies

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