제품특징
Advantages
· More Colonies, Ready Sooner!
- Cloning efficiency를
향상 시켜 빠른 colony 형성을 도와 줍니다.
· Robust
and Consistent Cloning.
- 다양한 배양 시스템 혹은 cell line에
걸쳐 유사한 성능을 보여 줍니다.
· Enhanced
Survival
- 다양한 seeding count혹은
electroporation 같은 높은 스트레스를 발생 시키는 실험 후 plating에 도움을 줍니다.
· Straight
to Single Cell
- Single-cell 계대 배양의
적응 단계를 건너 뛸 수 있도록 도와 줍니다.
Colony
Efficiency를
위한 제품의 비교
Figure
1. CloneR™ and CloneR™2 Supplements Improve
Cloning Efficiency and Colony Size
hPSCs display a considerable increase in
cloning efficiency when cloned using (B) CloneR™ compared to using (A) Y-27632
compound. (C) CloneR™2 further improves cloning efficiency and increases colony
size when compared to either Y-27632 compound or CloneR™. Shown are examples of
H9 hESCs in 10-cm dishes, plated at 200 cells per dish (~4 cells/cm²) in mTeSR™
Plus on Vitronectin XF™.
CloneR™과 CloneR™2의
비교
Figure
2. Use of CloneR™2 Enables Improved Cloning Efficiency and Larger Colonies
When Compared to Use of CloneR™
(A) Representative images of 200 cells (H9 cell line) in
12-well plates grown in mTeSR™1 on Vitronectin XF™ at day 8 after seeding.Three hES (H1, H7 and H9) and 5 hiPS (WLS-1C, STiPS-F016, STiPS-M001,
STiPS-R038 and STiPS-B004) cell lines were seeded at clonal density (50
cells/cm²) on Vitronectin XF™, in mTeSR™1 supplemented with CloneR™ or
CloneR™2. mTeSR™1 supplemented with CloneR™2 increases (B) cloning efficiency
and (C) colony size of hPSCs when compared with mTeSR™1 supplemented with CloneR™.
Each data point in (B) represents an average of 3 technical replicates, with at
least 7 biological replicates (n) per cell line.
Figure
3. CloneR™2 Improves Clonal Generation Following Single Cell Deposition
Single-cell deposition is the gold standard of cloning;
it relies on a FACS-based method that typically results in low cloning
efficiency. Clones generated in mTeSR™ Plus supplemented with CloneR™2
demonstrate significantly improved (E) cloning efficiency and (B, D) colony
size when compared to clones generated in mTeSR™ Plus supplemented with
CloneR™. (A, C) Representative images of H9 hESCs cultured for eight days
plated on Vitronectin XF™. Across four cell lines tested, CloneR™ and CloneR™2
had an average cloning efficiency of 21.2 ± 5.6% and 38.6 ± 6.0%, respectively,
with at least 3 biological replicates per cell line. * denotes p < 0.05; ***
denotes p < 0.001 by unpaired t-tests.
Y-27632와 CloneR™2의
비교
Figure
4. CloneR™2 Improves Seeding Efficiency at High Density
CloneR™2 improves single-cell seeding efficiency when
used as a supplement in media for the first 24 hours of culture, compared to
using Y-27632 as a supplement. 5.0x10⁵ cells were seeded in 12-well plates coated
with Corning® Matrigel® in mTeSR™ Plus
supplemented with CloneR™2 or Y-27632. Cultures were analyzed 24
hours post-seeding. The use of CloneR™2 resulted in an average seeding
efficiency of 98.2 ± 12.8% compared to the use of Y-27632, which resulted in an
average seeding efficiency of 81.9 ± 15.8%, across all cell lines (n = 3
replicates per line).
Figure
5. hPSCs Plated in CloneR™2 Show Increased Expansion
When used as a seeding supplement during single-cell
passaging, CloneR™2 improves cell expansion when compared to using Y-27632. 3.0x10⁴
cells were seeded in 12-well plates coated with Corning® Matrigel® in mTeSR™
Plus supplemented with CloneR™2 or Y-27632. After 24 hours, the cultures were
maintained in complete media (without a cloning supplement) and analyzed on day
5. CloneR™2 resulted in an average expansion of 49.1 ± 10.4 compared to
Y-27632, which resulted in a lower average expansion of 21.1 ± 8.2, across all
cell lines (n = 3 replicates per line).
Colony에 스트레스가 높은 시험을 진행 한 후의 증식 효율
Figure
6. CloneR™2 Improves Recovery of hPSCs Following Electroporation
CloneR™2 can also be used as a survival supplement in
gene-editing workflows that require electroporation. Four cell lines were
electroporated, then plated in mTeSR™1 and mTeSR™ Plus containing Y-27632,
CloneR™, or CloneR™2. Cultures were maintained in complete TeSR™ media (without
cloning supplement) after 24 hours and analyzed after 5 days. In all 4 cell
lines, (panels A-D) CloneR™2 dramatically improved cell survival and expansion
when used as a supplement in the first 24 hours immediately following
electroporation compared to both Y-27632 and CloneR™ (n = 2 replicates per cell
line).
Figure
7. CloneR™2 Improves Post-Thaw Recovery of hPSCs
Thawing cryopreserved cells can result in low expansion
or loss of the culture within the first passage. Using CloneR™2 as a seeding
supplement within the first 24 hours of thawing cells ameliorates this effect,
improving post-thaw recovery of hPSCs. Three cell lines were frozen as single
cells, then thawed into mTeSR™ Plus containing Y-27632 or CloneR™2 on
Matrigel®. Cultures were maintained in complete mTeSR™ Plus (without cloning
supplement) after 24 hours, and analyzed on day 4 or day 5. CloneR™2 improves
the fold-expansion across all cell lines tested when compared to Y-27632, with at
least 7 replicates (n) per cell line.
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