Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support
Present cell culture medium supplements, in most cases based on animal sera, are not fully satisfactory especially for the in vitro expansion of cells intended for human cell therapy. This paper refers to (i) an heparin-free human platelet lysate (PL) devoid of serum or plasma components (v-PL) and...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2017-11-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/article/10.3389/fbioe.2017.00066/full |
| _version_ | 1828897076669317120 |
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| author | Anita Muraglia Van Thi Nguyen Marta Nardini Massimo Mogni Domenico Coviello Beatrice Dozin Paolo Strada Ilaria Baldelli Matteo Formica Ranieri Cancedda Ranieri Cancedda Maddalena Mastrogiacomo |
| author_facet | Anita Muraglia Van Thi Nguyen Marta Nardini Massimo Mogni Domenico Coviello Beatrice Dozin Paolo Strada Ilaria Baldelli Matteo Formica Ranieri Cancedda Ranieri Cancedda Maddalena Mastrogiacomo |
| author_sort | Anita Muraglia |
| collection | DOAJ |
| description | Present cell culture medium supplements, in most cases based on animal sera, are not fully satisfactory especially for the in vitro expansion of cells intended for human cell therapy. This paper refers to (i) an heparin-free human platelet lysate (PL) devoid of serum or plasma components (v-PL) and (ii) an heparin-free human serum derived from plasma devoid of PL components (Pl-s) and to their use as single components or in combination in primary or cell line cultures. Human mesenchymal stem cells (MSC) primary cultures were obtained from adipose tissue, bone marrow, and umbilical cord. Human chondrocytes were obtained from articular cartilage biopsies. In general, MSC expanded in the presence of Pl-s alone showed a low or no proliferation in comparison to cells grown with the combination of Pl-s and v-PL. Confluent, growth-arrested cells, either human MSC or human articular chondrocytes, treated with v-PL resumed proliferation, whereas control cultures, not supplemented with v-PL, remained quiescent and did not proliferate. Interestingly, signal transduction pathways distinctive of proliferation were activated also in cells treated with v-PL in the absence of serum, when cell proliferation did not occur, indicating that v-PL could induce the cell re-entry in the cell cycle (cell commitment), but the presence of serum proteins was an absolute requirement for cell proliferation to happen. Indeed, Pl-s alone supported cell growth in constitutively activated cell lines (U-937, HeLa, HaCaT, and V-79) regardless of the co-presence of v-PL. Plasma- and plasma-derived serum were equally able to sustain cell proliferation although, for cells cultured in adhesion, the Pl-s was more efficient than the plasma from which it was derived. In conclusion, the cells expanded in the presence of the new additives maintained their differentiation potential and did not show alterations in their karyotype. |
| first_indexed | 2024-12-13T14:50:29Z |
| format | Article |
| id | doaj.art-6b610c56b5764dfb92c1a39de7201d33 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-12-13T14:50:29Z |
| publishDate | 2017-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-6b610c56b5764dfb92c1a39de7201d332022-12-21T23:41:21ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852017-11-01510.3389/fbioe.2017.00066286684Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation SupportAnita Muraglia0Van Thi Nguyen1Marta Nardini2Massimo Mogni3Domenico Coviello4Beatrice Dozin5Paolo Strada6Ilaria Baldelli7Matteo Formica8Ranieri Cancedda9Ranieri Cancedda10Maddalena Mastrogiacomo11Biorigen Srl, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyHuman Genetics Laboratory, E.O. Ospedali Galliera, Genoa, ItalyHuman Genetics Laboratory, E.O. Ospedali Galliera, Genoa, ItalyClinical Epidemiology, Ospedale Policlinico San Martino, Genoa, ItalyTransfusion Center, Ospedale Policlinico San Martino, Genoa, ItalyPlastic and Reconstructive Surgery, Ospedale Policlinico San Martino, Genoa, ItalyOrthopedic, Traumatology and Vertebral Surgery, Ospedale Policlinico San Martino, Genoa, ItalyBiorigen Srl, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyPresent cell culture medium supplements, in most cases based on animal sera, are not fully satisfactory especially for the in vitro expansion of cells intended for human cell therapy. This paper refers to (i) an heparin-free human platelet lysate (PL) devoid of serum or plasma components (v-PL) and (ii) an heparin-free human serum derived from plasma devoid of PL components (Pl-s) and to their use as single components or in combination in primary or cell line cultures. Human mesenchymal stem cells (MSC) primary cultures were obtained from adipose tissue, bone marrow, and umbilical cord. Human chondrocytes were obtained from articular cartilage biopsies. In general, MSC expanded in the presence of Pl-s alone showed a low or no proliferation in comparison to cells grown with the combination of Pl-s and v-PL. Confluent, growth-arrested cells, either human MSC or human articular chondrocytes, treated with v-PL resumed proliferation, whereas control cultures, not supplemented with v-PL, remained quiescent and did not proliferate. Interestingly, signal transduction pathways distinctive of proliferation were activated also in cells treated with v-PL in the absence of serum, when cell proliferation did not occur, indicating that v-PL could induce the cell re-entry in the cell cycle (cell commitment), but the presence of serum proteins was an absolute requirement for cell proliferation to happen. Indeed, Pl-s alone supported cell growth in constitutively activated cell lines (U-937, HeLa, HaCaT, and V-79) regardless of the co-presence of v-PL. Plasma- and plasma-derived serum were equally able to sustain cell proliferation although, for cells cultured in adhesion, the Pl-s was more efficient than the plasma from which it was derived. In conclusion, the cells expanded in the presence of the new additives maintained their differentiation potential and did not show alterations in their karyotype.http://journal.frontiersin.org/article/10.3389/fbioe.2017.00066/fullplatelet lysateplatelet factorsstem cell proliferationcell therapycell line culture |
| spellingShingle | Anita Muraglia Van Thi Nguyen Marta Nardini Massimo Mogni Domenico Coviello Beatrice Dozin Paolo Strada Ilaria Baldelli Matteo Formica Ranieri Cancedda Ranieri Cancedda Maddalena Mastrogiacomo Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support Frontiers in Bioengineering and Biotechnology platelet lysate platelet factors stem cell proliferation cell therapy cell line culture |
| title | Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support |
| title_full | Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support |
| title_fullStr | Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support |
| title_full_unstemmed | Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support |
| title_short | Culture Medium Supplements Derived from Human Platelet and Plasma: Cell Commitment and Proliferation Support |
| title_sort | culture medium supplements derived from human platelet and plasma cell commitment and proliferation support |
| topic | platelet lysate platelet factors stem cell proliferation cell therapy cell line culture |
| url | http://journal.frontiersin.org/article/10.3389/fbioe.2017.00066/full |
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