Feeder-dependent culture proven more reliable than FBS
Gibco KnockOut Serum Replacement (SR) – Multi-Species is a serum-free supplement that supports the growth of pluripotent stem cells (PSCs) cultured on fibroblast feeder cells. Designed to directly replace serum in existing protocols, this research-use product uses the same KnockOut Serum Replacement (KSR) formulation that has been trusted by many for almost 20 years in over 2,000 publications.
KnockOut SR – Multi-Species offers:
- A more defined formulation for feeder-dependent culture of mouse, human, and other mammalian PSCs
- More compact colony morphology compared to serum (Figure 1)
- Better maintenance of undifferentiated PSCs (Figure 2)
- Stable price and supply
Figure 1. PSC morphology in cultures grown using KnockOut SR – Multi-Species vs. FBS. H9 human embryonic stem cells (ESCs) were cultured on mouse embryonic fibroblasts (MEFs) with 20% fetal bovine serum (FBS) or 20% KnockOut SR – Multi-Species. 129S6 mouse ESCs were cultured on MEFs using 15% FBS or 15% KnockOut SR – Multi-Species. The human ESC colonies were thicker and more compact when cultured with KnockOut SR – Multi-Species than with FBS. The mouse ESC colonies cultured with KnockOut SR – Multi-Species were dome-shaped and more compact than with FBS.
Figure 2. PSC marker expression in cultures grown with KnockOut SR – Multi-Species vs. FBS. H9 human ESCs and 129S6 mouse ESCs were grown on MEFs under the same conditions as described in Figure 1. Human ESCs grown using KnockOut SR – Multi-Species exhibit uniformly high Oct4 (red) and Sox2 (green) expression, while those grown using FBS are heterogeneous and show more faint Sox2 staining. Mouse ESCs grown using KnockOut SR – Multi-Species also show higher and more homogeneous expression of Oct4 than those cultured in FBS. DAPI (blue) was used as a nuclear stain. Both cultures showed tighter, more desirable colony morphology when grown with KnockOut SR – Multi-Species.
Human PSCs already in culture can be passaged into medium containing KnockOut SR – Multi-Species using standard passaging protocols.
Transitioning mouse PSCs from FBS to KnockOut SR – Multi-Species
Mouse PSCs can thawed or passaged directly into medium containing KnockOut SR – Multi-Species using standard thawing or passaging protocols.
Table 1. Comparison of growth characteristics of human and mouse cells grown in serum and in KSR – Multi-Species.
|Species||Serum||KSR – Multi-Species||References|
|Colony morphology||Human||Less compact||More compact||H. Koivisto, 2004; M. Amit, 2000|
|Colony morphology||Mouse||Flatter & less compact||Dome-shaped & more compact||R. Blelloch, 2007; T. Davies, 2012|
|Growth rate||Human||Slower||Faster||H. Koivisto, 2004; J. Inzunza, 2005; H. Skottman, 2006|
|Unwanted differentiation||Human and mouse||High||Low||J. Inzunza, 2005; H. Koivisto, 2004; H. Skottman, 2006; P. Hong-mei, 2006; C. Denning, 2006; G. Guo, 2016|
|Efficiency of ESC derivation||Human and mouse||Low||Higher||P. Hong-mei, 2006; T. Davies, 2012; J. Cheng, 2004; V. Bryja, 2006|
|Clonal efficiency||Human and mouse||Low||Higher||M. Amit, 2000; J. Cheng, 2004|
|Reprogramming||Mouse||Slower||Faster, selectively supports PSCs||R. Blelloch, 2007; X. Zhao, 2010; H. Hirai, 2012|