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细胞培养基及添加物

TeSR™-E8™3D

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

概要
技术资料
数据及文献

概要

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.

技术资料

Document Type 产品名称 Catalog # Lot # 语言
Product Information Sheet TeSR™-E8™3D 03990 All English
Manual TeSR™-E8™3D 03990 All English
Safety Data Sheet 1 TeSR™-E8™3D 03990 All English
Safety Data Sheet 2 TeSR™-E8™3D 03990 All English
Safety Data Sheet 3 TeSR™-E8™3D 03990 All English

数据及文献

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.