Biological activity of a standardized freeze-dried platelet derivative to be used as cell culture medium supplement

Anita Muraglia, Chiara Ottonello, Raffaele Spanò, Beatrice Dozin, Paolo Strada, Michele Grandizio, Ranieri Cancedda, Maddalena Mastrogiacomo

Research output: Contribution to journalArticle

Abstract

Serum of animal origin and in particular fetal bovine serum is the most commonly utilized cell culture medium additive for in vitro cell growth and differentiation. However, several major concerns have been raised by the scientific community regarding the use of animal sera for human cell-based culture applications. Among the possible alternatives to the animal serum, platelet-derived compounds have been proposed since more than 10 years. Nevertheless, the high degree of variability between the different platelet preparations, and the lack of standardized manufacturing and quality control procedures, made difficult to reach a consensus on the applicability of this novel cell culture medium supplement. In this study, we describe the preparation of a standardized platelet-rich plasma (PRP) derivative obtained starting from human-certified buffy coat samples with a defined platelet concentration and following protocols including also freeze-drying, gamma irradiation and biological activity testing prior the product release as cell culture medium additive. Biological activity testing of the different preparations was done by determining the capability of the different PRP preparations to sustain human bone marrow mesenchymal stem cell (MSC) clone formation and proliferation. Taking advantage of a developed MSC in vitro clonogenicity test, we also determined biological activity and stability of the freeze-dried gamma-sterilized PRP preparations after their storage for different times and at different temperatures. The PRP effects on cell proliferation were determined both on primary cell cultures established from different tissues and on a cell line. Results were compared with those obtained in "traditional" parallel control cultures performed in the presence of bovine serum [10% fetal calf serum (FCS)]. Compared to FCS, the PRP addition to the culture medium increased the MSC colony number and average size. In primary cell cultures and in cell line cultures, the PRP promoted cell proliferation also in conditions where the FCS had not a proliferation stimulating effect due to either the nature of the cells and the tissue of origin (such as human articular chondrocytes from elderly patients) or to the critical low density cell seeding (such as for HeLa cells). In summary, the standardized PRP formulation would provide an "off-The-shelf" product to be used for the selection and expansion of several cell types also in critical cell culture conditions.

Original languageEnglish
Pages (from-to)211-220
Number of pages10
JournalPlatelets
Volume25
Issue number3
DOIs
Publication statusPublished - 2014

Fingerprint

Platelet-Rich Plasma
Culture Media
Blood Platelets
Cell Culture Techniques
Serum
Mesenchymal Stromal Cells
Primary Cell Culture
Cell Count
Cell Proliferation
Cell Line
Freeze Drying
Chondrocytes
Plasma Cells
HeLa Cells
Quality Control
Cell Differentiation
Clone Cells
Joints
Bone Marrow
Temperature

Keywords

  • Cell culture supplement
  • Lyophilized platelet-rich plasma
  • Storage temperature

ASJC Scopus subject areas

  • Hematology
  • Medicine(all)

Cite this

Biological activity of a standardized freeze-dried platelet derivative to be used as cell culture medium supplement. / Muraglia, Anita; Ottonello, Chiara; Spanò, Raffaele; Dozin, Beatrice; Strada, Paolo; Grandizio, Michele; Cancedda, Ranieri; Mastrogiacomo, Maddalena.

In: Platelets, Vol. 25, No. 3, 2014, p. 211-220.

Research output: Contribution to journalArticle

Muraglia, A, Ottonello, C, Spanò, R, Dozin, B, Strada, P, Grandizio, M, Cancedda, R & Mastrogiacomo, M 2014, 'Biological activity of a standardized freeze-dried platelet derivative to be used as cell culture medium supplement', Platelets, vol. 25, no. 3, pp. 211-220. https://doi.org/10.3109/09537104.2013.803529
Muraglia, Anita ; Ottonello, Chiara ; Spanò, Raffaele ; Dozin, Beatrice ; Strada, Paolo ; Grandizio, Michele ; Cancedda, Ranieri ; Mastrogiacomo, Maddalena. / Biological activity of a standardized freeze-dried platelet derivative to be used as cell culture medium supplement. In: Platelets. 2014 ; Vol. 25, No. 3. pp. 211-220.
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AU - Mastrogiacomo, Maddalena

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