Home › Cell Biology › The use of SC1 (Pluripotin) to Support mESC Self-renewal in the Absence of LIF
Cell Biology JoVE (Open Access) Citable · DOI

The use of SC1 (Pluripotin) to Support mESC Self-renewal in the Absence of LIF

DOI: 10.3791/1550-v
What you'll learn
  • ✓ Maintain mouse ES cell self-renewal without LIF using SC1
  • ✓ Assess pluripotency markers via immunocytochemistry
  • ✓ Evaluate SC1 efficacy through morphological and molecular analysis
Protocol

SC1 functions through dual inhibition of Ras- GAP and ERK1. We tested the function of SC1 in supporting mouse ES cell self-renewal in the absence of LIF and showed that SC1 is able to maintain self-renewal of mouse ES cell cultures.

Difficulty
intermediate
Total time
~5–7 days (culture maintenance and marker assessment)
Model organism
Mouse embryonic stem cells (mESC)
Biosafety
BSL-1

Steps

1
Understand SC1 mechanism and LIF-independent self-renewal

Learn how SC1 functions through dual inhibition of Ras-GAP and ERK1 to support mESC self-renewal without leukemia inhibitory factor (LIF).

▶ 00:08
2
Maintain mouse ES cell cultures with SC1

Culture and passage mESCs in the presence of SC1 to sustain self-renewal capacity over multiple passages without exogenous LIF supplementation.

▶ 00:51
3
Perform immunocytochemistry for pluripotency markers

Use immunofluorescence staining to detect and visualize pluripotency-associated markers in SC1-treated mESC cultures to confirm maintenance of stemness.

▶ 03:11
4
Examine and interpret representative SC1-treated images

Analyze representative microscopy images of mESCs grown in SC1 to assess morphology and marker expression patterns indicative of successful self-renewal.

▶ 04:54
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