| Cell Type |
Culture conditions |
Time-point for sample collection |
Viable cell count and viability at the time of sample collection |
Sample processing and storage |
Number of total proteins identified |
Known as secreted among all identified proteins (%) |
Potential factor(s) affecting outcome. |
| CHO-K1 [4] |
Cells were grown in in-house serum- and animal component-free chemically-defined media in bioreactor. |
2-, 3-, 4- and 5-days of culture |
Viable cell counts: ~ 1.8, 3.8, 7.4 and 9.5×106respectively
Viability: >95%. |
Supernatant was collected, centrifuged at 1000 rpm for 15 min and stored at -80°C. Spent media was then filter-purified (0.22 µM) and proteins were concentrated using methanol/chloroform precipitation. |
2512 |
11.54% |
High proportion of intracellular- and non-secretory proteins (~ 88.46%) among secretome possibly due to on-going cell death during harvest or mechanical damage by the bioreactor impellors and presence of cell-debris and micro vesicles in the samples. |
| CHO-S and DG44 [5] |
Cells were grown in chemically defined and serum-free CD-FortiCHO and CD-DG44 medium respectively in suspension culture. |
3-days of culture |
Viable Cell Counts: 100x106cells/ml
Viability: 98% |
- Spent-media was filter-purified (0.2 µM).
- Concentrated using 5 kDa molecular weight cut-offs by ultracentrifugation at 4°C. Protein sample were stored at -80°C. |
325 |
100% |
Selective enrichment of secretory proteins using metabolic labeling resulted in improved efficiency for identifying secreted-proteins only. |
| CHOK1SV[6] |
Cells were grown in serum-free CD-CHO media in suspension culture (fed-batch mini-bioreactors). |
7-, 11- and 15-days of culture |
Two bioreactors with ~ 72% viability and two with ~ 57% |
Spent-media were clarified by centrifugation at 900 rpm for 5 min and stored at -80°C. |
84 |
No information in the article |
High proportion of intracellular- and non-secretory proteins could be expected among secretome due to on-going cell-death and presence of cell-debris and micro vesicles in the samples. |
| CHO-K1 [28] |
Cells were grown in serum-free SFM4CHO media in suspension culture (shake flasks). |
5th-day of culture |
Viable cell count: 5-7 × 106 cells/mL
Viability: 97-99% |
Spent media was centrifuged at 180 ×g for 10 min, and stored at -80°C. Media was concentrated using 10 kDa molecular weight cut-offs. |
178 |
28% |
High proportion of intracellular- and non-secretory proteins (72%) among secretome possibly due to on-going cell death and presence of cell-debris and micro vesicles in the samples. |
| CHO-K1 [30] |
Cells were grown in serum-supplemented F-12K media till confluence and then shifted to serum-free F-12K media for 12hrs after washing cells with sterile PBS. |
80% confluency |
No information in the article |
- No information regarding centrifugation for removal of cell debris.
- Spent-media was collected and concentrated using 10 kDa molecular weight cut-offs. |
1977 |
No information in the article |
12 fractions were generated from the sample to increase secretome coverage. However, high proportion of intracellular- and non-secretory proteins could be expected among secretome due to high cell-death and presence of cell-debris and microvesicles in the samples. |
