Table 4 Immuno-phenotypic analysis performed and CD Marker Expression
Device/ System used (Author) | Type of immuno-phenotypic analysis of cell subtypes | Terminology for uncultured, freshly isolated cells | Stage of cell processing | Positive cell CD marker expression (%) | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Mesenchymal stem cell markers *CD markers observed in Pericytes as well | Endothelial cell, pericyte and haematopoetic markers | ||||||||||||||
CD 13 | CD 29 | CD 44* | CD 73 | CD 90* | CD 105* | CD 146* | CD 31 | CD 34 | CD 45 | CD 68 | Other | ||||
Adinizer (Copcu et al. [18]) | Flow Cytometry | Stromal cells/ Nuclear cells | Immediately after device use (minimally manipulated) | As per methods- Proportions of CD45 negative cells were analysed in CD34−CD146+ and CD34+CD146−CD90+ (deemed as regenerative perivascular cells), and CD34+CD146+ as endothelial cells. However, percentages not specifically reported in results. | |||||||||||
No control | |||||||||||||||
Passage in culture following device (extensively manipulated) | |||||||||||||||
Adiprep (Dragoo et al. [27]) | Flow Cytometry | SVF Cells | Immediately after device use (minimally manipulated) | 56.5 | 72.0 | 60.4 | 65.2 | 33.4 | 80.3 | ||||||
No control | |||||||||||||||
Passage in culture following device (extensively manipulated) | 94.3 | 96.6 | 97.0 | ||||||||||||
Fastem (Domenis et al. [24]) | Flow Cytometry | SVF Cells | Immediately after device use (minimally manipulated) | 50-60 | CD34+CD45-CD31- 10-20 | ||||||||||
Control- ‘modified’ Coleman’s procedure (centrifugation) | 0-10 | CD34+CD45-CD31- 20-30 | |||||||||||||
Passage in culture following device (extensively manipulated) | |||||||||||||||
Fastem and MyStem (Gentile et al. [29]) | Not done | SVF Nucleated Cells | |||||||||||||
Hy-Tissue SVF (Busato et al. [12]) | Flow Cytometry | Free nucleated SVF cells | Immediately after device use (minimally manipulated) | 7.61 | 6.28 | 2.6 | 9.91 | 5.5 | 3.5 | CD116 0.7 | |||||
Control- enzymatic digestion using 0.1% collagenase type I at 37 °C for 45min followed by centrifugation at 400G for 10min. | 10.1 | 9.98 | 3.67 | ||||||||||||
Passage in culture following device (extensively manipulated) | 90 | >90 | >70 | 60 | 90 | ||||||||||
Lipocube Nano (Cohen et al. [17]) | Flow Cytometry | SVF Cells | Immediately after device use (minimally manipulated) | 42.0 | 53 | 55.8 | 53.2 | 18.8 | |||||||
No control | |||||||||||||||
Passage in culture following device (extensively manipulated) | |||||||||||||||
Lipocube SVF (Tiryaki et al. [66]) | Flow Cytometry | Nucleated SVF Cells | Immediately after device use (minimally manipulated) | 21.5 | 6.16 | 11.4 | 9.0 | ||||||||
Control- enzymatic digestion using GMP grade collagenase NB6 at a concentration of 0.1 U/ml at 37 °C for 30min followed by centrifugation at 400G for 10min. Then washed with PBS solution and centrifuged at 300G for 5min. | 6.93 | 3.44 | 5.88 | 3.06 | |||||||||||
Passage in culture following device (extensively manipulated) | |||||||||||||||
Lipogems (Vezzani et al. [75]) | Flow Cytometry | SVF Nucleated Cells | Immediately after device use (minimally manipulated) | CD146+CD34 33.5 CD34+CD146 5.46 | |||||||||||
No control | CD146+CD34 8.39 CD34+CD146 51.5 | ||||||||||||||
Passage in culture following device (extensively manipulated) | CD14, CD31, CD40 ligand (CD154) significantly more abundant than when compared to control. | ||||||||||||||
Lull pgm (Morselli et al. [42]) | Not done | SVF Cells | |||||||||||||
MyStem (Cicione et al. [16]) | Flow Cytometry | Lipoaspirate fluid cells | Immediately after device use (minimally manipulated) | <0.1 | 1-1.5 | <0.1 | 0.5-1 | <0.5 | <1 | ||||||
Control- centrifugation ‘as previously described’ | <0.1 | 1.5-2 | <0.1 | 1 | <0.5 | <0.5 | |||||||||
Passage in culture following device (extensively manipulated) | 93 | 98 | 95 | 96 | |||||||||||
MyStem (Tarallo et al. [65]) | Flow Cytometry | Freshly isolated LAF Cells | Immediately after device use (minimally manipulated) | 0-10 | 75 | 0-10 | 0-10 | 20 | CD31 30 | ||||||
No control | |||||||||||||||
Passage in culture following device (extensively manipulated) | All culture-expanded cells displayed an ASC-like immunophenotype: CD105+, CD73+, CD90+, CD45- and CD34-CD31. | ||||||||||||||
Puregraft (Streit et al. [63]) | Direct Immunofluorescence | SVF Cells | Immediately after device use (minimally manipulated) | Analysed adhesive properties to determine stem cell nature. All adherent cells were positive for CD90 and CD105 and negative for CD31 and CD45 antigens (stem cell marker). Numbers not specified. | |||||||||||
Control 1- aliquot was left at 37°C for 20min under the action of gravity (decantation). Control 2- aliquot centrifuged at 1200G for 3 min. | |||||||||||||||
Passage in culture following device (extensively manipulated) | |||||||||||||||
Rigenera (Dai Pre et al. [20]) | Flow Cytometry | Total cells | Immediately after device use (minimally manipulated) | 3.12 | 4.98 | CD44/CD90 30.4 CD73/CD105 16.6 CD73/29 27.8 | |||||||||
Control- enzymatic digestion using 0.1% collagenase type I at 37 °C for 45min in Hank’s Balanced Salt Solution (HBSS) and 2% bovine serum albumin followed by centrifugation at 3000 rpm for 7min. | 76.7 | 7.32 | CD44/CD90 48.1 CD73/CD105 54.3 CD73/29 62 | ||||||||||||
Passage in culture following device (extensively manipulated) | Expression of the typical mesenchymal stem cell markers (CD105, CD90, CD73, CD44, and CD29) and the hematopoietic markers (CD45 and CD34) was preserved through culture passages. | ||||||||||||||
Transpose RT (Winnier et al. [77]) | Not done | Adipose-derived regenerative cells | |||||||||||||
Tulip Nanotransfer (Cohen et al. [17]) | Flow Cytometry | SVF Cells | Immediately after device use (minimally manipulated) | 18.3 | 50 | 42.1 | 24.1 | 7.9 | |||||||
No control | |||||||||||||||
Passage in culture following device (extensively manipulated) | |||||||||||||||
Tulip Nanotransfer (Sese et al. [61]) | Not done | Nanofat cells | |||||||||||||