Blood was collected into heparin-coated tubes. Collected blood was diluted with phosphate buffered saline (PBS) and centrifuged through a Ficoll gradient (Catalogue number, 17-1440-03, GE Healthcare, Little Chalfont, Buckinghamshire, UK) for 30 minutes at 850 ×g. PBMCs were collected, transferred to a new tube, washed, and resuspended in StemSpan medium (9805, STEMCELL Technologies, Vancouver, British Columbia, Canada) supplemented with CC110 cytokine cocktail (8697, STEMCELL Technologies). Cells were maintained for 5 days at 5% CO2, at 37°C, before use.
Cc110 cytokine cocktail
Manufactured by STEMCELL
Sourced in Canada
The CC110 cytokine cocktail is a specialized cell culture media supplement formulated to support the expansion and maintenance of human CD34+ hematopoietic stem and progenitor cells. The cocktail contains a proprietary combination of cytokines and growth factors that promote cell proliferation and differentiation.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using cc110 cytokine cocktail
1
Isolation and Culture of Human PBMCs
2
Isolation and culture of peripheral blood mononuclear cells from systemic sclerosis patients
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Blood was collected into heparin tubes, diluted with phosphate-buffered saline (PBS), and centrifuged through a Ficoll gradient (GE Healthcare) for 30 min at 850 g. The PBMCs were washed, resuspended in StemSpan medium (STEMCELL Technologies) supplemented with CC110 cytokine cocktail (STEMCELL Technologies), and cultured for 5 days at 37℃ in 5% CO2 before being reprogrammed. PBMCs were collected from five SSc patients subjects, and the clinical information of the patients is shown in Table 1 .
Clinical information of systemic sclerosis patients
| No | Sex | Age range | Disease duration, years | Skin involvement | ILD | PAH | GI involvement | ANA | Anti-centromere | Anti-scl70 |
|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 2 | 40–49 | 20 | Limited | Yes, NSIP | No | GERD | 1:80, nucleolar | Negative | Negative |
| 2 | 1 | 40–49 | 13 | Diffuse | Yes, NSIP + UIP | Yes | Recurrent paralytic ileus | 1:1280, speckled | Negative | Negative |
| 3 | 1 | 50–59 | 2 | Diffuse | Yes, UIP | No | GERD | 1:1280, cytoplasmic | Negative | Positive |
| 4 | 2 | 60–69 | 5 | Diffuse | Yes, NSIP | No | GERD | 1:80, speckled | Negative | Negative |
| 5 | 1 | 50–59 | 16 | Diffuse | No | No | GERD, scleroderma esophagus | 1:640, speckled | Negative | Negative |
1, Female; 2, Male; ILD, interstitial lung disease; PAH, pulmonary artery hypertension; ANA, antinuclear antibody; NSIP, nonspecific interstitial pneumonia; UIP, usual interstitial pneumonia; GERD, gastroesophageal reflux disease
3
Isolation and Culture of Cord Blood Mononuclear Cells
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CBMCs were directly obtained from the Cord Blood Bank of the Seoul St. Mary’s Hospital. Cord blood was diluted with phosphate-buffered saline (PBS) and centrifuged through Ficoll gradient (GE Healthcare, Little Chalfont, UK) for 30 minutes at 850 × g. CBMCs were collected, washed, and frozen. Prior to being used, frozen CBMCs were thawed and resuspended in StemSpan medium (Stemcell Technologies, Vancouver, BC, Canada) supplemented with CC110 cytokine cocktail (Stemcell). Cells were maintained for 5 days at 5% CO2, 37 °C before reprogramming.
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