搜索结果: 'methocult media formulations for human hematopoietic cells serum containing'

科学海报

Optimized Media and Workflow for the Expansion of Human Pluripotent Stem Cells as Aggregates in Suspension Cultures

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Conference：

ISSCR 2019

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发布日期: 07/05/2019

New Tools for the Ex Vivo Expansion of Human Hematopoietic Stem and Progenitor Cells

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New Tools for the Ex Vivo Expansion of Human Hematopoietic Stem and Progenitor Cells

发布日期: 03/09/2015

S. S. De Ravin et al. (APR 2016) Nature biotechnology 34 4 424--9

Targeted gene addition in human CD34(+) hematopoietic cells for correction of X-linked chronic granulomatous disease.

Gene therapy with genetically modified human CD34(+) hematopoietic stem and progenitor cells (HSPCs) may be safer using targeted integration (TI) of transgenes into a genomic 'safe harbor' site rather than random viral integration. We demonstrate that temporally optimized delivery of zinc finger nuclease mRNA via electroporation and adeno-associated virus (AAV) 6 delivery of donor constructs in human HSPCs approaches clinically relevant levels of TI into the AAVS1 safe harbor locus. Up to 58{\%} Venus(+) HSPCs with 6-16{\%} human cell marking were observed following engraftment into mice. In HSPCs from patients with X-linked chronic granulomatous disease (X-CGD),caused by mutations in the gp91phox subunit of the NADPH oxidase,TI of a gp91phox transgene into AAVS1 resulted in ∼15{\%} gp91phox expression and increased NADPH oxidase activity in ex vivo-derived neutrophils. In mice transplanted with corrected HSPCs,4-11{\%} of human cells in the bone marrow expressed gp91phox. This method for TI into AAVS1 may be broadly applicable to correction of other monogenic diseases. View Publication

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09600

09650

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StemSpan™ SFEM

J. Yen et al. (NOV 2018) Scientific reports 8 1 16304

TRIAMF: A New Method for Delivery of Cas9 Ribonucleoprotein Complex to Human Hematopoietic Stem Cells.

CRISPR/Cas9 mediated gene editing of patient-derived hematopoietic stem and progenitor cells (HSPCs) ex vivo followed by autologous transplantation of the edited HSPCs back to the patient can provide a potential cure for monogenic blood disorders such as $\beta$-hemoglobinopathies. One challenge for this strategy is efficient delivery of the ribonucleoprotein (RNP) complex,consisting of purified Cas9 protein and guide RNA,into HSPCs. Because $\beta$-hemoglobinopathies are most prevalent in developing countries,it is desirable to have a reliable,efficient,easy-to-use and cost effective delivery method. With this goal in mind,we developed TRansmembrane Internalization Assisted by Membrane Filtration (TRIAMF),a new method to quickly and effectively deliver RNPs into HSPCs by passing a RNP and cell mixture through a filter membrane. We achieved robust gene editing in HSPCs using TRIAMF and demonstrated that the multilineage colony forming capacities and the competence for engraftment in immunocompromised mice of HSPCs were preserved post TRIAMF treatment. TRIAMF is a custom designed system using inexpensive components and has the capacity to process HSPCs at clinical scale. View Publication

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22001

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22011

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STEMvision™ 人脐带血7-天CFU分析包

STEMvision™ 彩色人脐带血14-天CFU分析包

STEMvision™ 彩色人骨髓14-天CFU分析包

STEMvision™ 彩色人动员外周血14-天CFU分析包

STEMvision™ 小鼠总CFU分析包

STEMvision™ 小鼠髓系CFU分析包

STEMvision™ 小鼠红系CFU分析包

STEMvision™ 小鼠CFU分析包（髓系和红系）

实验方案

Transitioning from Feeder-Free Media to mTeSR™ Plus for Human Pluripotent Stem Cell Culture

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研究方向：

干细胞生物学

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发布日期: 02/10/2020

Ali N et al. (APR 2009) Blood 113 16 3690--5

Forward RNAi screens in primary human hematopoietic stem/progenitor cells.

The mechanisms regulating key fate decisions such as self-renewal and differentiation in hematopoietic stem and progenitor cells (HSPC) remain poorly understood. We report here a screening strategy developed to assess modulators of human hematopoiesis using a lentiviral short hairpin RNA (shRNA) library transduced into cord blood-derived stem/progenitor cells. To screen for modifiers of self-renewal/differentiation,we used the limited persistence of HSPCs under ex vivo culture conditions as a baseline for functional selection of shRNAs conferring enhanced maintenance or expansion of the stem/progenitor potential. This approach enables complex,pooled screens in large numbers of cells. Functional selection identified novel specific gene targets (exostoses 1) or shRNA constructs capable of altering human hematopoietic progenitor differentiation or stem cell expansion,respectively,thereby demonstrating the potential of this forward screening approach in primary human stem cell populations. View Publication

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09600

09650

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StemSpan™ SFEM

Bruserud &O et al. (MAY 2003) Leukemia research 27 5 455--64

In vitro culture of human acute lymphoblastic leukemia (ALL) cells in serum-free media; a comparison of native ALL blasts, ALL cell lines and virus-transformed B cell lines.

The aim of this study was to standardize in vitro culture conditions for human acute lymphoblastic leukemia (ALL) cells. The cells were cultured in medium containing 10% fetal calf serum (FCS) and in the four serum-free media X-vivo 10,X-vivo 15,X-vivo 20 and Stem Span. Native ALL blasts could proliferate in all four serum-free media,but the strongest responses were usually observed with Stem Span. Native leukemia blasts were also cultured in the presence of various single cytokines or cytokine combinations. The highest proliferation was usually observed in the presence of Flt3-Ligand (Flt3-L) when single cytokines were examined,and these responses could be further increased especially by combining Flt3-L with interleukin 3 (IL3),IL7 or stem cell factor (SCF). Proliferation could also be increased when ALL blasts were cultured in the presence of two commercially available fibroblast cell lines (Hs27 and HFL1). Based on these results we suggest that in vitro culture conditions for native human ALL blasts can be standardized by using serum-free culture media supplemented with exogenous Flt3-L+IL3+SCF,and the use of accessory cells can also be standardized by using well-characterized fibroblast cell lines. Detectable ALL blast proliferation can then be observed for most patients. Our experimental model can thereby be used for in vitro evaluation of possible antileukemic treatment strategies,and it will then allow comparison of experimental results between different studies. View Publication

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Otsuji TG et al. (MAY 2014) Stem Cell Reports 2 5 734--745

A 3D sphere culture system containing functional polymers for large-scale human pluripotent stem cell production

Utilizing human pluripotent stem cells (hPSCs) in cell-based therapy and drug discovery requires large-scale cell production. However,scaling up conventional adherent cultures presents challenges of maintaining a uniform high quality at low cost. In this regard,suspension cultures are a viable alternative,because they are scalable and do not require adhesion surfaces. 3D culture systems such as bioreactors can be exploited for large-scale production. However,the limitations of current suspension culture methods include spontaneous fusion between cell aggregates and suboptimal passaging methods by dissociation and reaggregation. 3D culture systems that dynamically stir carrier beads or cell aggregates should be refined to reduce shearing forces that damage hPSCs. Here,we report a simple 3D sphere culture system that incorporates mechanical passaging and functional polymers. This setup resolves major problems associated with suspension culture methods and dynamic stirring systems and may be optimal for applications involving large-scale hPSC production. ?? 2014 The Authors. View Publication

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05850

05857

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mTeSR™1

技术手册

Human Colony-Forming Unit (CFU) Assays Using MethoCult™

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00217

00217UK

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00602

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04436

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04534

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04437

04447

00215US.4

00215CA.3

00215GER.3

0

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MethoCult™ SF H4436

MethoCult™ H4034 Optimum 入门试剂盒

MethoCult™ H4100

MethoCult™ H4230

MethoCult™ SF H4236

MethoCult™ H4431

MethoCult™ H4434 Classic

MethoCult™ H4434 Classic 套装

MethoCult™ H4531

MethoCult™ H4535 Enriched，不含EPO

MethoCult™ SF H4536

MethoCult™ H4534 Classic 无 EPO 入门试剂盒

含有10% 牛血清白蛋白（BSA）的 Iscove's MDM

MethoCult™ 不含EPO的H4035 Optimum

MethoCult™ H4330

MethoCult™ H4034 Optimum

MethoCult™ H4435 Enriched

MethoCult™ H4534 Classic（不含 EPO）

MethoCult™ Express

造血祖细胞检测标准化培训课程

科学海报

Efficient Differentiation of Human Pluripotent Stem Cells to Hematopoietic Progenitor Cells in Serum-Free Culture Conditions

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Conference：

ISEH 2016

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05310

产品名：

STEMdiff™ 造血试剂盒

L. Truszkowski et al. (Sep 2025) Open Research Europe 4 2

Refined and benchmarked homemade media for cost-effective, weekend-free human pluripotent stem cell culture

Cost-effective,practical,and reproducible culture of human pluripotent stem cells (hPSCs) is required for basic and translational research. Basal 8 (B8) has emerged as a cost-effective solution for weekend-free and chemically-defined hPSC culture. However,the requirement to home-produce some recombinant growth factors for B8 can hinder access and reproducibility. Moreover,we found the published B8 formulation suboptimal in widely-used normoxic hPSC culture. Lastly,the performance of B8 in functional applications such as genome editing or organoid differentiation required systematic evaluation. We formulated B8 with commercially available,growth factors and adjusted its composition to support normoxic culture of WTC11 human induced pluripotent stem cell line. We compared this formulation (B8+) with commercial Essential 8 (cE8) and a home-made,weekend-free E8 formulation (hE8). We measured pluripotency marker expression and cell cycle by flow cytometry,and investigated the transcriptional profiles by bulk and single-cell RNA sequencing. We further assessed genomic stability,genome editing efficiency,single-cell cloning,and differentiation in both monolayer and organoids. Finally,we validated key findings using male (H1) and female (H9) human embryonic stem cells. hE8 performed comparably to cE8 across most functional assays and cell lines. In contrast,cells in B8+ displayed higher NANOG expression and improved genome editing efficiency. At the same time,B8+ led to gene expression changes indicative of marked lineage priming,reflected in altered morphology and differential response to some differentiation protocols. Both weekend-free media resulted in a modest transcriptional shift towards a less metabolically active state,consistent with intermittent media starvation. Homemade weekend-free media can provide a cost-effective alternative to commercial formulations. hE8,integrating some features of B8 while resembling cE8,emerges as a robust and practical option with limited compromises. B8+,though advantageous in some contexts,warrants caution due to lineage priming effects that may impact differentiation outcomes. View Publication

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05230

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STEMdiff™ 三胚层分化试剂盒

技术公告

StemSpan™ Media and Supplements for the Expansion and Differentiation of Erythroid Progenitor Cells

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产品号#：

02692

09600

09650

09605

09655

09955

产品名：

StemSpan™红系扩增添加物 (100X)

StemSpan™ SFEM

StemSpan™ SFEM II

发布日期: 02/01/2016

搜索结果: 'methocult media formulations for human hematopoietic cells serum containing'

Targeted gene addition in human CD34(+) hematopoietic cells for correction of X-linked chronic granulomatous disease.

TRIAMF: A New Method for Delivery of Cas9 Ribonucleoprotein Complex to Human Hematopoietic Stem Cells.

Forward RNAi screens in primary human hematopoietic stem/progenitor cells.

In vitro culture of human acute lymphoblastic leukemia (ALL) cells in serum-free media; a comparison of native ALL blasts, ALL cell lines and virus-transformed B cell lines.

A 3D sphere culture system containing functional polymers for large-scale human pluripotent stem cell production

Refined and benchmarked homemade media for cost-effective, weekend-free human pluripotent stem cell culture

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