TeSR™-E8™3D

Animal component-free suspension culture medium for human ES and iPS cells
TeSR™-E8™3D

Animal component-free suspension culture medium for human ES and iPS cells

1 Kit
Catalog # 03990
Required Products
  1. AggreWell™ Rinsing Solution
    Anti-Adherence Rinsing Solution

    Rinsing solution for cultureware to prevent cell adhesion

  2. Gentle Cell Dissociation Reagent
    Gentle Cell Dissociation Reagent

    cGMP, enzyme-free cell dissociation reagent

  3. 37μm Reversible Strainers|27215,27250
    Reversible Strainers

    Reversible strainer for filtration of single cells and isolation of cellular aggregates, including embryoid bodies and spheroids

  4. Y-27632 (Dihydrochloride)
    Y-27632

    RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2

Overview

TeSR™-E8™3D is an animal component-free culture medium developed for the expansion and scale-up of undifferentiated human embryonic stem (ES) and induced pluripotent stem (iPS) cells as aggregates in 3D suspension culture.

Based on TeSR™-E8™, TeSR™-E8™3D is a low-protein medium with an optimized protocol for scaling up human ES and iPS cell production in suspension culture. The novel fed-batch workflow saves time and media; daily feeds replenish nutrients and eliminate the need for medium exchanges on non-passaging days.

TeSR™-E8™3D is made with the highest level of quality and care. It contains only the essential components required for scale-up of ES and iPS cells. The TeSR™-E8™3D system is compatible with cells previously cultured in mTeSR™1 or TeSR™-E8™ on Corning® Matrigel® hESC-Qualified Matrix (Corning Catalog #354277) or Vitronectin XF™ (Catalog #07180, a matrix developed and manufactured by Nucleus Biologics), and is compatible with a range of suspension culture vessels.
Advantages
• OPTIMIZED: Part of the TeSR™ family, mTeSR™3D and TeSR™-E8™3D are optimized for hPSC suspension culture
• STREAMLINED WORKFLOW: Fed-batch strategy provides a simplified culture system
• SIMPLIFIED CULTURE SYSTEM: Serum-free culture system with no microcarriers or external matrix required
• SCALE UP: Easily produce up to 1 x 10^9 hPSCs in as few as 2-3 weeks
• COST EFFECTIVE: Significant cost savings in both media and labor
Components
  • TeSR™-E8™3D (Catalog #03990)
    • TeSR™-E8™3D Seed Basal Medium, 480 mL
    • TeSR™-E8™3D Seed 25X Supplement, 20 mL
    • TeSR™-E8™3D Feed Supplement, 20 mL
Subtype
Specialized Media
Cell Type
Pluripotent Stem Cells
Species
Human
Application
Cell Culture, Expansion
Brand
TeSR
Area of Interest
Stem Cell Biology
Formulation
Animal Component-Free

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
TeSR™-E8™3D
Catalog #
03990
Lot #
All
Language
English
Document Type
Manual
Product Name
TeSR™-E8™3D
Catalog #
03990
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
TeSR™-E8™3D
Catalog #
03990
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
TeSR™-E8™3D
Catalog #
03990
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
TeSR™-E8™3D
Catalog #
03990
Lot #
All
Language
English

Educational Materials (7)

Webinar
Nature Research Round Table: Parkinson's Disease Therapy with Human Embryonic Stem Cells
24:46
Nature Research Round Table: Parkinson's Disease Therapy with Human Embryonic Stem Cells
Webinar
Nature Research Round Table: Standards for Pluripotent Stem Cell Banking
13:02
Nature Research Round Table: Standards for Pluripotent Stem Cell Banking
Webinar
Nature Research Round Table: Human Pluripotent Stem Cell Lines as Disease Models
17:30
Nature Research Round Table: Human Pluripotent Stem Cell Lines as Disease Models
Webinar
Nature Research Round Table: Defining and Maintaining Pluripotency & hPSC Line Registration and Banking - Panel Discussion
33:47
Nature Research Round Table: Defining and Maintaining Pluripotency & hPSC Line Registration and Banking - Panel Discussion
Webinar
Nature Research Round Table: hPSC Lines for Cell Therapies - Panel Discussion
29:03
Nature Research Round Table: hPSC Lines for Cell Therapies - Panel Discussion
Webinar
Nature Research Round Table: Retinal Cell Therapy Using Human Embryonic Stem Cells
19:35
Nature Research Round Table: Retinal Cell Therapy Using Human Embryonic Stem Cells
Webinar
Nature Research Round Table: HLA Typing Considerations for Human Pluripotent Stem Cell Banking
13:01
Nature Research Round Table: HLA Typing Considerations for Human Pluripotent Stem Cell Banking

Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications

Data

Expansion of human pluripotent stem cells (ESC and iPSC) in TeSR™-E8™3D suspension culture

Figure 1. Human pluripotent stem cell (hPSC) growth in TeSR™-E8™3D

TeSR™-E8™3D supports expansion of hPSCs over multiple passages in aggregate suspension culture. Shown are (A) cumulative fold expansion and (B) daily fold expansion in human ES cell lines (H1 and H7) and human iPS cell lines (WLS-1C and STiPS-F016) over 5 passages. Error bars are +/- SD, n=3.

Viability of human pluripotent stem cells (ESC and iPSC) cells in TeSR™-E8™3D suspension culture over time

Figure 2. Viability of hPSCs in TeSR™-E8™3D

hPSCs cultured in TeSR™-E8™3D maintain high viability over multiple passages. Shown are end-of-passage viabilities for human ES cell lines (H1 and H7) and human iPS cell lines (WLS-1C and STiPS-F016) over 5 passages. Error bars are +/- SD, n=3.

Human ES and iPS cell aggregates in TeSR™-E8™3D suspension culture shown at 20x and 40x magnification

Figure 3. Morphology of hPSC aggregates in TeSR™-E8™3D

hPSCs depict morphology characteristic of hPSC aggregates in suspension culture including mostly spherical shape, clear edges (not smooth or shiny), even color, and dimpled or pock-marking appearance. Aggregates should remain under 400 µm in size.

Glucose and lactate levels (mM) in human ES and iPS cultures over time in TeSR™-E8™3D

Figure 4. Metabolic Profile of hPSCs during culture in TeSR™-E8™3D

(A) Glucose concentration in TeSR™-E8™3D is maintained at optimal levels throughout the culture period. Dotted line represents the glucose level in TeSR™-E8™3D Complete Seed Medium. Error bars are +/- SD, n = 3, and (B) end-of-passage lactate concentration remains below 15 mM throughout culture in TeSR™-E8™3D. Error bars are +/- SD, n = 3

Percentage of OCT-4 and TRA-1-60 positive cells in human ES and iPS TeSR™-E8™3D suspension cultures

Figure 5. Markers of undifferentiated hPSCs measured in TeSR™-E8™3D cultures.

Shown are (A) OCT-4 expression and (B) TRA-1-60 expression for multiple hPSC lines after 5 passages in TeSR™-E8™3D. Error bars are +/- SD, n = 3.

Expression of lineage-specific markers measured in ectoderm, mesoderm, and endoderm differentiated from human ES and iPS cells cultured in TeSR™-E8™3D suspension culture.

Figure 6. Trilineage differentiation of hPSCs cultured in TeSR™-E8™3D.

Trilineage differentiation potential to was assessed using the STEMdiff™ Trilineage Differentiation Kit. hPSCs exhibit equivalent or better differentiation to (A) ectoderm, (B) mesoderm, and (C) endoderm compared to 2D controls, after 5 passages in TeSR™-E8™3D. Error bars are +/- SD, n = 3.

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