BACKGROUND:Biobanks have evolved from simple repositories into complex research infrastructures that require robust governance frameworks to ensure traceability, data integrity, and operational reliability. ISO 20387 provides internationally harmonized accreditation standards for biobanking; however, empirical evidence evaluating its organizational impact remains limited. METHODS:We conducted a longitudinal observational study of a hospital-based biobank between 2019 and 2025. ISO 20387 accreditation obtained in 2022 was considered the structural intervention point in the analysis. Governance and quality indicators were evaluated across the sample lifecycle, including labeling error rate, metadata completeness, traceability completeness, nonconformity rate per 100 aliquots, on-time processing, and Corrective Action and Preventive Action (CAPA) closure time. RESULTS:Operational activity expanded substantially during the study period, with annual aliquot production increasing from 2142 aliquots in 2019 to a peak of 7668 in 2022. Governance indicators improved progressively during the pre-accreditation phase and stabilized following accreditation. Metadata completeness increased from 74% to 86% before accreditation to 99%-100% afterward, while traceability completeness rose from 71%-73% to 98%-99%. Nonconformity rates declined from 2.03-2.43 per 100 aliquots before accreditation to 0.75-0.90 afterward. On-time processing improved from 87%-93% to 98.5%-99%, and median CAPA closure time decreased from 53-63 to 39-42 days. CONCLUSIONS:ISO 20387 accreditation was associated with structural stabilization of governance processes in a hospital-based biobank undergoing substantial operational expansion. These findings suggest that accreditation can act as a catalyst for governance maturation, helping biobanking infrastructures maintain operational control and quality performance while scaling activity.

BACKGROUND:ISO 20387:2018 is the first international standard specifically designed for biobanks, defining requirements for competence, impartiality, and operational consistency. In 2022, the Multi-Specialistic Biobank of the Azienda Ospedaliero-Universitaria Pisana became the first biobank in Italy to achieve ISO 20387 accreditation, setting a national benchmark for public-sector quality governance. APPROACH:The accreditation pathway followed a structured 12-month plan combining gap analysis, document harmonization, competence development, and internal audits. A multidisciplinary quality working group redesigned the quality management system and aligned managerial and operational processes with ISO 20387 requirements. Fourteen quality documents were revised or newly developed to standardize workflows, strengthen traceability, and embed risk-based principles throughout the biobank's activities. OUTCOMES:ISO 20387 accreditation was granted on July 14, 2022. Subsequent surveillance audits in 2023, 2024, and 2025 confirmed sustained compliance and progressive improvements, including expanded internal audits, enhanced risk assessment, digital competence tracking, and strengthened Corrective and Preventive Action effectiveness. Persistent challenges-including infrastructural constraints, limited information technology support, and delays in software modifications-reflected structural limitations typical of public health care settings. SIGNIFICANCE:This article provides an integrated account of ISO 20387 implementation and long-term maintenance in a public hospital biobank. The Pisa experience offers a replicable model for institutions operating under similar constraints, demonstrating how technical rigor, coordinated governance, and a shared quality culture can sustain accreditation and advance national biobanking excellence.

Background and aims: This paper aims to study an alternative solution to hormonal replacement therapy in specific groups of patients who underwent thyroidectomy during childhood or adulthood. After cryopreservation, thyroid autotransplantation could be an alternative solution which would allow us to use the ability of the thyroid tissue of producing hormones according to the physiological needs of the body. Materials and methods: A feasibility study about the effects of the most modern cryopreservation techniques on the structural and functional integrity of the follicular cells of the thyroid tissue has been carried out. Patients who could benefit from the treatment have been found for both autotransplant techniques. Additionally, a literature review has been conducted. Results: The histological analysis has shown that cryopreservation does not alter the original architecture, and the culture examination that cell viability is successfully preserved. Moreover, both thyroid autotransplantation studies on animals and those on humans that were found in the literature have shown good results regarding the viability and functionality of the transplant. Conclusions: The viability of cryopreserved thyroid tissue found in this study is encouraging. Further studies to evaluate the levels of FT3, FT4 and thyroglobulin in thyroid tissue after cryopreservation are needed to verify that the secretory properties of the thyrocytes have been maintained intact. Furthermore, autotransplanted cases found in the literature do not have a long-term follow-up.

BACKGROUND Carotid artery disease is highly prevalent and a main cause of ischemic stroke and vascular dementia. There is a paucity of information on predictors of serious vascular events. Besides percentage diameter stenosis, international guidelines also recommend the evaluation of qualitative characteristics of carotid artery disease as a guide to treatment, but with no agreement on which qualitative features to assess. This inadequate knowledge leads to a poor ability to identify patients at risk, dispersion of medical resources, and unproven use of expensive and resource-consuming techniques, such as magnetic resonance imaging, positron emission tomography, and computed tomography. OBJECTIVES The Carotid Artery Multimodality imaging Prognostic (CAMP) study will: prospectively determine the best predictors of silent and overt ischemic stroke and vascular dementia in patients with asymptomatic subcritical carotid artery disease by identifying the noninvasive diagnostic features of the 'vulnerable carotid plaque'; assess whether 'smart' use of low-cost diagnostic methods such as ultrasound-based evaluations may yield at least the same level of prospective information as more expensive techniques. STUDY DESIGN We will compare the prognostic/predictive value of all proposed techniques with regard to silent or clinically manifest ischemic stroke and vascular dementia. The study will include ≥300 patients with asymptomatic, unilateral, intermediate degree (40-60% diameter) common or internal carotid artery stenosis detected at carotid ultrasound, with a 2-year follow-up. The study design has been registered on Clinicaltrial.gov on December 17, 2020 (ID number NCT04679727).

Departmental Section of Biobanks (SOD Biobanche) of the Azienda Ospedaliera Universitaria Pisana, consists of a Tissue and Cell institute, the Heart Valve Bank, and a biorepository, the Regional Biologic Archive. A clean room consisting of three laboratories (1 class C and 2 class B) represents a core facility for some of the processes and projects of the biobank. The Regional Biological Archive is a biobank which currently carries out as an institutional task the storage of serum, plasma and non-viable cells of multiorgan, multi-tissue donors and donors of corneas, as well as solid organ transplant recipients of the Tuscany Region, providing the possibility of reviewing and re-evaluating even at a distance of years the donor/recipient pairs. The mission of SOD Biobanche is to act as a transversal service platform in the field of biobanking of biological material. In this broader perspective the structure is open to new projects concerning biobanking, some of which are already in the activation phase, such as a bank for urogenital tissues, a biobank of tumors of the musculoskeletal system and an advanced therapy protocol for the use of partially osteoinduced expanded mesenchymal stem cells.

BACKGROUND:Bone Marrow MSCs are an appealing source for several cell-based therapies. Many bioreactors, as the Quantum Cell Expansion System, have been developed to generate a large number of MSCs under Good Manufacturing Practice conditions by using Human Platelet Lysate (HPL). Previously we isolated in the human bone marrow a novel cell population, named Mesodermal Progenitor Cells (MPCs), which we identified as precursors of MSCs. MPCs could represent an important cell source for regenerative medicine applications. As HPL gives rise to a homogeneus MSC population, limiting the harvesting of other cell types, in this study we investigated the efficacy of pooled human AB serum (ABS) to provide clinically relevant numbers of both MSCs and MPCs for regenerative medicine applications by using the Quantum System.METHODS:Bone marrow aspirates were obtained from healthy adult individuals undergoing routine total hip replacement surgery and used to generate primary cultures in the bioreactor. HPL and ABS were tested as supplements to culture medium. Morphological observations, cytofluorimetric analysis, lactate and glucose level assessment were performed.RESULTS:ABS gave rise to both heterogeneous MSC and MPC population. About 95% of cells cultured in HPL showed a fibroblast-like morphology and typical mesenchymal surface markers, but MPCs were scarcely represented.DISCUSSION:The use of ABS appeared to sustain a large scale MSC production, as well as the recovery of a subset of MPCs, and resulted a suitable alternative to HPL in the cell generation based on the Quantum System.

BACKGROUND AIMS:Mesenchymal stromal cells (MSCs) modulate the immune response and represent a potential treatment for inflammatory and autoimmune diseases. We hypothesized that this feature could be potentiated by co-administering anti-inflammatory cytokines. In this article, we asked whether engineering of Wharton Jelly-derived human MSCs (WJ-hMSCs) to express an anti-inflammatory cytokine increases cell immunomodulatory properties without altering their native features.METHODS:We used Epstein-Barr virus-derived interleukin-10 (vIL-10), which shares some immunosuppressive properties with human IL-10 but lacks immunostimulatory activity. Engineering was accomplished by transducing WJ-hMSCs with a self-inactivating feline immunodeficiency virus-derived vector co-expressing vIL-10 and herpes simplex virus type-1 thymidine kinase (TK). TK was added to allow future tracking of WJ-hMSC in vivo by positron electron tomography (PET).RESULTS:The results show that (i) expression of TK and/or vIL-10 does not change WJ-hMSC phenotypic and functional properties; (ii) vIL-10 is secreted, biologically active and enhances the immunosuppressing functions of WJ-hMSCs; (iii) v-IL10 and TK can be produced simultaneously by the same cells and do not interfere with each other.DISCUSSION:WJ-hMSCs engineered to secrete vIL-10 could be a powerful tool for adoptive cell therapy of immune-mediated diseases, and therefore, additional studies are warranted to confirm their efficacy in suitable animal disease models.

Dear Sir, It is well known that umbilical cord blood (UCB) is an alternative source of stem cells for haematopoietic transplantation, which can cure children and adults of malignant and non-malignant diseases, such as leukaemias, lymphomas, metabolic disorders, immunodeficiency states, haemoglobinopathies and bone marrow failure syndromes1–2. Cord blood processing and cryopreservation are very costly procedures. In order to reduce these costs several pre-birth parameters were suggested to be useful for identifying and selecting the potential best collections3 . A survey-based study was recently performed in Greece on a sample of Greek citizens, with high reproductive capacity. The results indicated that the majority of them (74%) were in favour of UCB storage and that the main source of information (45.5%) was the media, with an additional source provided by medical health care professionals (36.6%)4. However, most of the Greek population do not have adequate information about this precious resource and the authors believed that different campaigns regarding UCB storage and uses should be developed through the cooperation of the Ministry of Health and Social Solidarity, the National Blood Donation Centre and the National Transplant Organisation4. In accordance with the findings of the Greek study, a survey conducted by Screnci et al. at the “Sapienza” University of Rome on blood donors and pregnant women showed that a considerable proportion of the individuals analysed had some general, but minimal knowledge about UCB. Moreover, the level of knowledge regarding the proper and real usefulness was generally low: 31% of informed pregnant women gave a wrong answer. A very small proportion of them consulted the specific website section of the Ministry of Health concerning the appropriate use of stem cells from UCB5. Despite the theoretically positive intention of Greek respondents towards UCB storage, most did not proceed to do so. Among the parents who chose to store UCB, most preferred private storage, perhaps because of the misleading information that exists in Greece about public banks in contrast to private banks. The reason given for choosing private banks was that the UCB would be stored better. Moreover, private storage was encouraged by doctors, friends or relatives and by the belief that it guarantees the baby’s future4. A survey-based study was conducted from the 1st September 2013 to the 31st of January 2014, in the whole region of Tuscany. The aim of the survey was to explore the knowledge that pregnant women have about UCB stem cell donation and the quality of education from health care professionals. For this purpose anonymous, standardised printed or on-line questionnaires were provided. Printed questionnaires were given to 2,999 women (average age=31.91) who delivered in birth centres in 14 Hospitals, while on-line questionnaires were sent to gynaecologists and 706 midwives who are involved in raising awareness about donation and cord blood collection. The surveyed population was divided based on the birth centre of origin (Table I). Table I Distribution of the sample. Since only eight gynaecologists filled in the questionnaire, our attention focused on pregnant women and midwives. Among 2,999 mothers who were asked to fill in the questionnaire, 1,568 (52%) returned the completed form; 60% of the respondents had not had previous pregnancies. The women were asked whether they had received general information about UCB donation during pregnancy. A considerable portion of pregnant women (82%) were aware of the possibility of donating UCB, while 18% had not received any information. The most common sources of information came from midwives (41.90%), gynaecologists (17.16%), family doctors (1.4%) and other sources, such as friends (14.60%) and media (7.53%). Several women received information from more than one of these sources. The 706 midwives involved in this study were divided in different groups depending on how long they had worked. While 70.49% of them had not received specific training on cord blood donation during their degree course, 69.4% of the midwives had attended a post-degree course about cord blood donation. Overall, 68.31% of midwives claimed to promote cord blood donation; however, although they claimed to be competent enough (mean=6.4) about their own skill and knowledge, 95.63% of them believed that further specific cord blood training would be necessary. About three-quarters (72.60%) of midwives use printed leaflets to give information, while only 1.37% of them use verbal communication. The majority of midwives spend an average of 13 minutes to give information. The critical aspects that arose from the survey highlight some solutions that could be adopted in order to improve the quality of information provided to pregnant women. The lack of interest shown by gynaecologists is one aspect which limits the choice of donating UCB. For this reason further involvement of these professionals will be necessary in the future. Midwives who had been employed for longer did not receive specific training on UCB collection during their degree course, even if they showed greater participation in training courses, in comparison with their colleagues who had started work more recently. However, all midwives declared that specific training courses would be important to update their knowledge on UCB donation and to make colleagues aware of this topic. Moreover, the time spent by midwives to inform pregnant women is thought to be insufficient. Finally, we consider that divulgation of information by means of leaflets should be combined with verbal communication, which is known to be the best way of raising people’s awareness. We believe that in our regional reality Hospitals should invest more money and time in promoting UCB donation, mostly because our data show that 58% of midwives don’t usually provide with information about this topic. Finally, we believe that professionals and institutions should make an effort to provide updates, education and evidence-based information about UCB donation, focusing on both the usefulness and limitations of UCB stem cells.

Background: Equine amnion mesenchymal stem cells (EAMSCs) from amnion isolated aft er the foal birth represented an alternative source of easy collection of mesenchymal cells used in equine regenerative medicine.Materials and Methods: Th ese cells grown as two-dimensional (2-D) culture in alpha modifi ed minimum essential medium supplemented with epidermal growth factor were diff erentiated in adipogenic, chondrogenic, and osteogenic cells.Half a million cells as pellet were left in 15 ml tubes with the same diff erentiation media for 20 days.Aft er the pellets were collected, embedded in paraffi n for morphological study.Results: 2-D culture showed EAMSCs with an embryonic phenotype (C-kit+, CD105+, Oct-4+) and a diff erentiation potential in adipogenic, chondrogenic and osteogenic multipotent cells.By a reproducible method of three-dimensional (3-D) culture, at day 20 the Authors evidenced a formation of small aggregated spheroids gradually gathering.In cross sections, the surface of the spheroid evidenced fl attened cells embedded in a red matrix by Alizarin staining and occasionally a core of calcium precipitation.A network of apoptotic or necrotic cells in a not mineralized matrix was present into the center of nodules.Th e 3-D spheroids appeared larger (mean diameter of 605 ± 53 μm for gathering spheroids and 1486 ± 79 μm for spheroids already gathered) than those from standard monolayer cultures (mean diameter of 200 ± 73 μm).Conclusions: EAMSCs cultured in 3D method preserve their in vitro multipotent diff erentiation than adherent 2-D culture method.Th ese EAMSCs included in the extracellular matrix not mineralized at day 20 seem to be a good source of MSCs for tissue repair and regeneration in equine medicine.

Dear Sir, Due to the detrimental action of free radicals and reactive oxygen species (ROS), oxidative stress is involved in the pathogenesis of ageing and diseases such as inflammation, cancer, cardiovascular disorders and infections. Although this view remains still valid, during the last decade the understanding of oxidative stress has progressively changed and it has become clear that free radicals and reactive oxygen species also play a key role in cell biology and function. In fact, oxidation/reduction reactions are a primary mechanism for regulation of cell proliferation, death, and most notably, cell differentiation, which involves the function of several redox-sensitive molecular elements1. The main intracellular antioxidant of mammalian cells, the tripeptide glutathione (GSH), plays a crucial role in such functions, together with other less abundant low molecular weight thiols (LMWT); in fact, besides participating in detoxicant reactions catalysed by glutathione transferase and glutathione peroxidase, the binding of GSH to proteins (protein S-glutathionylation) is a potent modulator of the molecular elements involved in cell proliferation, differentiation and death, both inside the cytoplasm (e.g. the transcription factor nuclear factor-B), and on the cell surface (e.g. tumour necrosis factor-α receptor)1. The cell surface enzyme gamma-glutamyltransferase (GGT) has a peculiar role in determining the biological action of GSH, as it catalyses the hydrolysis of the tripeptide GSH (gamma glutamylcysteinylglycine) to glutamate and the dipeptide cysteinyl-glycine (cys-gly). This latter is a potent reductant which promotes pro-oxidant reactions through the reduction of oxygen to superoxide anion radical and the generation of thiol radical. GGT is, therefore, able to transform an antioxidant (GSH) into a potent pro-oxidant (cys-gly), provided that sufficient oxygen is present in the environment2. The isolation, characterisation, in vitro expansion and differentiation of human umbilical cord blood mesenchymal stem cells (hUCBMSC) are among the current aims of regenerative medicine, and achieving these goals is a pre-requisite for extensive use of this novel approach for the treatment of a number of human diseases; unfortunately, reliable procedures for efficient expansion and differentiation of hUCBMSC remain to be established, and oxidative stress caused by culture conditions might be a factor to take into account when devising adequate conditions for maintaining and differentiating stem cells in vitro3. It is well known that standard cell culture conditions, which involve the exposure of cells to an atmosphere of air containing 5% CO2, are far from the physiologic situation. In fact, while the atmospheric oxygen tension is 160 mmHg, it reaches only 40 mmHg in blood, and decreases to 18 mmHg in foetal blood. Cell cultures are thus exposed to a potent oxidative stress even in basal conditions, and the gap between physiological and cell culture conditions is far wider if considering foetal tissues, which are subjected to very low oxygen concentrations in vivo4. We decided to check whether decreasing oxygen tension in culture to very low levels, corresponding to those found in foetal tissues, is compatible with cell growth in vitro, whether it has appreciable effects on the status of intracellular and extracellular low-molecular weight thiols of cultured mesenchymal stem cells, and whether GGT acts as an oxygen-dependent trigger of oxidative stress in vitro. Thirty-four human umbilical cord blood samples were harvested from term or preterm deliveries at the time of birth with the mothers' consent. The mean volume of the cord blood units (CBU) was 113±12.3 mL without anticoagulant. Blood samples were processed within 24 hours of collection. The mononuclear cells were separated using Percoll (Amersham Biosciences, Uppsala, Sweden) and were suspended in culture medium (low-glucose Dulbecco's modified Eagle's medium; GIBCO) containing 20% foetal calf serum, 100 U/mL penicillin, 100 μg/mL streptomycin and 2 mM L-glutamine. Cells were then seeded at a density of 1×106 cells/cm2 in 25 cm2 culture flasks. The human

Plasma samples from human cord blood, and fetuses, newborns, and adults of different mammalians species were analyzed by gel-filtration chromatography, to ascertain whether gamma-glutamyltransferase (GGT) fractions reflect liver maturation. Human cord blood plasma showed higher b-, m-, and s-GGT fraction as compared to adult women. In rat and mouse fetuses and in newborns, b-GGT was the most abundant fraction. As in adult humans, in adult rats, mice, rabbits, sheep, and mini pigs, f-GGT was the most abundant fraction. GGT fractions are a common feature of all mammalian species tested. Their pattern changes seem to reflect liver postnatal maturation, function.

The amnion is a particular tissue whose cells show features of multipotent stem cells proposed for use in cellular therapy and regenerative medicine. From equine amnion collected after the foal birth we have isolated MSCs (mesenchymal stem cells), namely EAMSCs (equine amnion mesenchymal stem cells), from the mesoblastic layer. The cells were grown in α-MEM (α-modified minimum essential medium) and the effect of EGF (epidermal growth factor) supplementation was evaluated. To assess the growth kinetic of EAMSCs we have taken into account some parameters [PD (population doubling), fold increase and DT (doubling time)]. The differentiation in chondrogenic, adipogenic and osteogenic types of cells and their epitope expression by a cytofluorimetric study have been reported. EGF supplementation of the culture medium resulted in a significant increase in PD growth parameter and in the formation of bone nodules for the osteogenic differentiation. By immunohistochemistry the amnion tissue shows a positivity for the c-Kit (cluster tyrosine-protein kinase), CD105 and Oct-4 (octamer-binding transcription factor 4) antigens that confirmed the presence of MSCs with embryonic phenotype.

The biological significance of donor-specific microchimerism (DSM) in solid organ transplantation is unresolved. It has been reported both as a favourable feature, which may facilitate induction and maintenance of tolerance, and as a sign of graft-vs-host disease. Here, we applied a quantitative real-time PCR assay (qRT-PCR) to a selected series of kidney transplant recipients to measure the level of microchimerism in relation to allograft function and survival. DSM level was assessed by scoring the HLA-DRB1 locus in 54 patients (42 males, 12 females) with more than 2 years of follow-up after transplantation; 38 patients were considered to have stable renal function (SRF) and 16 had allograft dysfunction (AD). Among patients with AD, 12 (75%) showed detectable level of microchimerism, compared to 11 (29%) SRF patients (Odds Ratio 7.36, 95% CI 1.7-35.2; p<0.01). In addition, AD patients showed a higher mean donor genome equivalents (6.5×10(-5) vs. 2.4×10(-5); p<0.001). SRF patients were re-evaluated two years later; 2 out of 27 DSM negative vs. 2 out of 11 DSM positive had lost their transplanted organ. In conclusion, qRT-PCR applied to peripheral blood shows significant association between DSM and allograft dysfunction in kidney transplant patients.

BACKGROUND:. The fact that only a small percentage of cord blood units (CBU) stored are actually used for transplantation contributes to raising the already high costs of their processing and cryopreservation. The identification of predictors allowing the early identification of suitable CBU would allow a reduction of costs for the collection, storage and characterisation of CBU with insufficient volume or cell numbers. In our bank we have adopted a cut-off value for using CBU of 8 x 10(8) nucleated cells and a volume >or= 60 mL.MATERIALS AND METHODS:In 365 banked CBU, we evaluated the correlation between neonatal/gestational parameters and laboratory data used to assess their quality.RESULTS:Biparietal diameter (BPD) and abdominal circumference were significantly and positively correlated with CBU volume (r(2)=0.12, p=0.0011 and r(2)=0.092, p=0.0063, respectively). Receiver operating characteristic (ROC) analysis showed that both parameters can be used to identify CBU with insufficient volume (BPD: area under the curve 0.69, 95% CI=0.57-0.82, p=0.004; abdominal circumference: area under the curve 0.67, 95% CI=0.54-0.79, p<0.01). BPD and head circumference, but not abdominal circumference or femoral length, were positively correlated with white blood cell (WBC) count (r(2)=0.215, p=0.031, and r(2)=0.299, p=0.015, respectively). Abdominal circumference, but not BPD, head circumference or femoral length, was statistically significantly correlated with the number of CD34(+) cells in the CBU. Weight at birth and placental weight were positively correlated with WBC count, blood volume, CD34(+) cell count, total colony-forming units and burst-forming units.CONCLUSION:. Pre-birth assessment of BPD might allow the selection of donors who would yield CBU of sufficient volume and WBC count and avoid the costs of collecting, transferring, storing and analysing CBU with a high probability of resulting unsuitable for transplantation.

Stem cells from extra-embryonic sources can be obtained by non-invasive procedures. We have standardized a method for the expansion of equine umbilical cord-derived matrix cells (EUCMCs) for potential therapy. EUCMCs were isolated from the umbilical cord of five mares immediately after delivery. For expansion, cells were grown in alpha-MEM and MSCBM. Moreover, to measure the effect of growth factor supplementation, epidermal growth factor (EGF) was added to alpha-MEM. alpha-MEM and MSCBM media performed similarly in terms of population doubling and CFU number value. EGF supplementation of alpha-MEM determined a significant increase of the population doubling value. EGF supplementation did not affect the adipogenic and chondrogenic differentiation while bone nodule sizes an increased with the osteogenic protocol. Both alpha-MEM and MSCBM can be used to cultivate EUCMCs. alpha-MEM supplemented with EGF might represent an advantage for EUCMCs expansion. The results could be useful in choosing the culture medium since alpha-MEM is more cost-effective than MSCBM.

Background - Rabbits provide an excellent model for many animal and human diseases, such as cardiovascular diseases, for the development of new vaccines in wound healing management and in the field of tissue engineering of tendon, cartilage, bone and skin. The study presented herein aims to investigate the biological properties of bone marrow rabbit MSCs cultured in different conditions, in order to provide a basis for their clinical applications in veterinary medicine. Findings - MSCs were isolated from 5 New Zealand rabbits. Fold increase, CFU number, doubling time, differentiation ability and immunophenotype were analyzed. With the plating density of 10 cells/cm 2 the fold increase was significantly lower with DMEM-20%FCS and MSCs growth was significantly higher with αMEM-hEGF. The highest clonogenic ability was found at 100 cell/cm 2 with MSCBM and at 10 cell/cm 2 with M199. Both at 10 and 100 cells/cm 2 , in αMEM medium, the highest CFU increase was obtained by adding bFGF. Supplementing culture media with 10%FCS-10%HS determined a significant increase of CFU. Conclusion - Our data suggest that different progenitor cells with differential sensitivity to media, sera and growth factors exist and the choice of culture conditions has to be carefully considered for MSC management.