Recent advances in Stem Cell culture: Encapsulated Stembeads growth factors

Activin A StemBeads tebu-bio

During my daily discussions with researchers involved in in vitro iPS and Embryonic Stem cell cultures, I am always amazed by the passion that drives them. However, the more I go in depth with them in their protocols to optimize them, the more I realize that “time and money” are very tricky points when cultivating these cells. This can be summarized by a simple question: How can I optimise my media to cultivate Stem cells with stable and cost-effective Stem Cell Factors?

In other words, what are the most economical recent advances in Stem Cell research allowing high quality culture without spending my week-ends feeding my Stem cells ?

Encapsulated Stem cell growth factors

Recently, a new technology based on “Stabilized” Stem cell Factors has been developed. These growth factors are encapsulated in biocompatible microspheres (Polyesters of Glycolic and Lactic Acids or PLGA beads). These “beads” allow sustained release of the Stem cell Factors into your media of choice, thus creating a more stable environment for your cells.

Optimal FGF-2 dependent stem cell culture – FGF2 StemBeads

StemBeads FGF2 steadily release growth factor into the media, thus enabling you to reduce the feeding frequency of your stem cell cultures to bi-weekly.

FGF-2 StemBeads users report the following advantages:

  • Significant savings on media costs – media changes are reduced by 67%
  • Better culture quality – reduced spontaneous differentiation
  • No protocol change – 100% compatibility with established media & culture conditions
  • Used with leading commercial media, it can save us up to 35% per week in media cost
  • Used with standard DMEM/KSR media, it can save up to 15%
FGF-2 StemBeads by tebu-bio vs soluble FGF-2

Culture with FGF-2 StemBeads and biweekly feeding produces a more undifferentiated hESC culture than traditional daily feeding. hESCs grown one month on MEF feeders exhibit a similar FACS stem cell profile when fed with FGF-2 StemBeads in medium every third day compared to soluble FGF-2 in medium daily. NANOG expression was significantly increased and the diffeerentiation markers SOX17 while Brachyury were significantly reduced, by qRT-PCR (left panel). Normal Karyotypes for hESCs were assessed before expansion and then after 1 month of expansion in either soluble FGF-2 or FGF-2 StemBeads, and neither showed abnormalities. Immunostaining of month old cultures shows similar appearance of colonies and expression of the pluripotency markers OCT4 and NANOG in both conditions, Scale = 50 microns. Source: StemCulture and Lotz et al. (20123, http://dx.doi.org/10.1371/journal.pone.0056289). Note: Similar results obtained with hESCs grown on Matrigel (BD).

FGF-2 StemBeads by tebu-bio on E12 Mouse NPCs

FGF-2 StemBeads produce a more undi#erentiated, mouse and human neural stem cell culture. Mouse NSCs grown for one week in FGF-2 StemBeads show increased progenitor cells (Nestin+) and decreased neuronal differentiation (TUJ1+) compared to no FGF-2 and soluble FGF-2, (scale bars = 50 microns). Source: StemCulture and Lotz et al. (20123, http://dx.doi.org/10.1371/journal.pone.0056289). Note: Similar results obtained with hESCs grown on Matrigel (BD).

A cost effective way to grow Activin-A dependent cell cultures – Activin A StemBeads

Activin-A is used sometimes to maintain pluripotent stem cells or to differentiate them to different lineages (ex. endoderm). While soluble Activin-A is degraded in culture after 12 hours of culture, StemBeads Activin-A releases a constant rate of Activin-A into your culture media. Data have shown that this Activin A release lasts for over 5 days, thus creating a more stable environment for Stem cell cultures.

Activin A users noted the following points:

  • Fewer media changes (as little as one media change in 5 days)
  • Significant savings compared to soluble Activin-A
  • Significant savings on media and less labor time
  • Easy-to-use : simply add Activin A StemBeads to usual media
Stability of StemBeads Activin A vs.  Soluble Activin A Stem Factor at tebu-bio
Sustained levels of Activin A StemBeads: Soluble Activin A (left) ot Activin A StemBeads (right) were added to culture media and left for 3 days. Media was collected and Activin A assessed every 24 hours by ELISA. Activin-A StemBeads remain stable over the time. Source: StemCulture.
Sox17 expression with StemBeads Activin A tebu-bio

Activin A StemBeads differentiates hESCs to Endodermal Lineage. hESCs were grown to 80-90% confluency in a KSR based media using FGF2 StemBeads. Media was refreshed and FGF-2 StemBeads was replaced by either soluble Activin-A or Activin A StemBeads. The Soluble Activin-A condition had the media changed once in 5 days, while the Activin A StemBeads condition was never changed afer initial addition. Activin A StemBead conditions yield to comparable SOX17+ cells with no media changes over 5 days. Source: StemCulture.

EGF dependent cell cultures – EGF StemBeads

These EGF StemBeads offer you the same reliability and flexibility as FGF2 and Activin A StemBeads. EGF is a growth factor that stimulates proliferation of various cell types and Stem cells. EGF StemBeads stabilize EGF leading to optimal cellular performance.

As these have only recently been released, I have not yet been able to collect user’s comments, but I have found some convincing data to show you that EGF StemBeads are genuinely efficient!

 

E14 and E18 Mouse cortical cells grown with EGF StemBeads tebu-bio

NSCs grown for one week in and EGF beads shows increased cell density over no EGF and Soluble EGF. (Blue: No EGF; Red: 10ng/ml Soluble EGF; Green: 10ng/ml EGF StemBeads). Source StemCell.

EGF StemBeads vs soluble EGF in IF staining of E14 and E18 Cortical cells tebu-bio

Mouse NSCs grown (1 week) with EGF StemBeads show increased progenitor cells (Nestin+) and Neurons (B-Tubulin-III) to no EGF and soluble EGF. Source: StemCulture.

Convinced that StemBead technology will save you time & money in your Stem Cell research?

To get started with StemBeads, contact our experts in cell culture, Stem cell and cell reprogramming approaches!

 

Written by Jean-François Têtu, PhD
Jean-François Têtu is Sales Manager at tebu-bio for the south of Europe and also for the UK and Eire. He also fervently defends and promotes anything from his home territory, the north of France.