The HALO® SPC-QC Potency Platform
The HALO® Stem and Progenitor Cell -
Quality Control (SPC-QC) Platform
The Colony-Forming Assay Alternative
The HALO® SPC-QC Platform is a special and logical extension application of our HALO® Research Platform. The rationale to develop the HALO® SPC-QC Kit was four-fold.
- To provide a rapid, standardized, non-subjective and validated quality control assay, these being characteristics that the traditional colony-forming assay has not attained.
- To demonstrate that cells processed for transplantation purposes exhibit proliferative (growth) potential.
- Predict long- and short-term engraftment.
- Prediction of long- and short-term balanced, multilineage reconstitution.
HemoGenix® now has 2 HALO® SPC-QC Platforms:- HALO®-96 MeC SPC-QC, which is similar to the traditional colony-forming assay in that it uses methyl cellulose, but does not require manual colony counting since it utilizes a very convenient, rapid, non-subjective and fully standardized ATP-based bioluminescence readout.
- HALO®-96 SEC (Suspension Expansion Culture) SPC-QC. This HALO® SPC-QC Platform does use methyl cellulose. The assay detects proliferation of the expanding cells in suspension culture. It is easier to use than HALO®-96 MeC because viscous methyl cellulose does not have to be dispensed. As a result, all procedures can be performed with normal pipettes. Results are obtained in only 5 days and the sensitivity of the assay is greater than the HALO®-96 MeC. Like HALO®-96 MeC, it is fast, non-subjective and fully standardized using an ATP-based bioluminescence readout.
Both HALO®-96 MeC and HALO®-96 SEC SPC-QC platforms can completely replace the traditional colony-forming assay because there is a direct statistical correlation between all 3 assay systems. The graph below shows that for human multipotential stem cells (CFC-GEMM) there is a correlation between HALO®-96MeC performed on day 7 and manual counting of colonies using the traditional colony-forming assay, performed on day 14.
The following 3-dimensional graph shows the relationship between the colony-forming assay, HALO®-96 MeC and HALO®-96 SEC.
There are now 2 alternative non-subjective and fully standardized assays to the colony-forming assay. From the table below, HALO®-96 SEC is more sensitive and and can be completed in less time than the HALO®-96 MeC. It is just up to you whether you prefer to use methyl cellulose or not. However, regardless of which HALO® Platform you use, you are ensuring the best possible information regarding the potency of the stem cell product.
| Colony-Forming Assay
| CAMEO™-96 STD
| HALO®-96 MeC SPC-QC
| HALO®-96 SEC SPC-QC |
| Assay detects: | Differentiation potential
| Proliferation and differentiation potential | Proliferation potential
| Proliferation potential
|
Type of assay:
| Differentiation
| Proliferation + differentiation
| Proliferation
| Proliferation
|
Type of culture:
| Methylcellulose
| Methylcellulose | Methylcellulose
| Suspension
|
Cell growth:
| Clonal
| Clonal | Clonal
| Expansion
|
Parameter measured:
| Cell colonies
| Cell colonies + intracellular ATP
| Intracellular ATP
| Intracellular ATP
|
Readout type:
| Manual
| Manual + Instrument-based
| Instrument-based
| Instrument-based
|
Readout:
| Colony number
| Colony number + bioluminescence | Bioluminescence
| Bioluminescence
|
| Subjectivity: | Subjective
| Non-subjective | Non-subjective
| Non-subjective
|
Calibration/ Standardization:
| None | External ATP with high and low controls
| External ATP with high and low controls
| External ATP with high and low controls
|
Format:
| 35mm Petri dishes
| 96-well plate | 96-well plate
| 96-well plate
|
Volume of assay:
| 1ml
| 100µl | 100µl
| 100µl
|
No. of replicates:
| 2-3
| 6 | 6 | 6 |
Sensitivity:
| 100-1,000 cells
| 25 cells (depends on purity) | 25 cells (depends on purity)
| 15-25 cells (depends on purity)
|
Incubation time:
| 14 days
| 14 days | 7 days
| 5 days
|
Processing / Readout time:
| None/~10min/plate
| ~1-2min/well / 15min/plate | 15 min/plate
| 15 min/plate
|
Throughput:
| Low
| Low | Medium
| Medium to High
|
Training:
| 6-12 months to count colonies
| 6-12 months to count colonies / 2 days
| 2 days
| 2 days
|
| Validation potential: | Extremely difficult | Yes | Yes
| Yes
|
Proficiency testing potential:
| None
| Yes | Yes
| Yes
|
