The Posterior Iliac Crest Outperforms the Anterior Iliac Crest When Obtaining Mesenchymal Stem Cells from Bone Marrow

Pierini, Michela; Bella, Claudia Di; Dozza, Barbara; Frisoni, Tommaso; Martella, Elisa; Bellotti, Chiara; Remondini, Daniel; Lucarelli, Enrico; Giannini, Sandro; Donati, Davide María

doi:10.2106/jbjs.l.00429

Cited by 95 publications

(67 citation statements)

References 3 publications

(1 reference statement)

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“…The local Ethical Committee approved the study and patients signed informed consent before the donation. MSC were isolated using gradient separation (Ficoll-Paque PREMIUM, density 1.073 g/mL; GE Healthcare, Uppsala, Sweden) and plastic adherence, as previously described (Pierini et al 2013). Briefly, after the gradient separation step, viable mononucleated cells (seeding density: 400,000 cells/cm 2 ) were seeded in complete medium composed of a-modified minimum essential medium (a-MEM; BioWhittaker, Lonza, Verviers, Belgium) supplemented with 20 % lot-selected fetal bovine serum (FBS; Lonza, Basel, Switzerland), 1 % GlutaMAX TM (Gibco, Life Technologies, Paisley, UK).…”

Section: Msc Isolation and Culturementioning

confidence: 99%

Long term morphological characterization of mesenchymal stromal cells 3D spheroids built with a rapid method based on entry-level equipment

et al. 2016

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Three-dimensional (3D) spheroids of mesenchymal stromal cells (MSC) have been demonstrated to improve a wide range of MSC features, such as multilineage potential, secretion of therapeutic factors, and resistance against hypoxic condition. Accordingly, they represent a promising tool in regenerative medicine for several biological and clinical applications. Many approaches have been proposed to generate MSC spheroids. They usually require specific generation systems, such as rotatory bioreactors or low-attachment plates, and each approach has its own disadvantages. Furthermore, an over-time analysis of morphological homogeneity and architectural stability of the spheroids generated is rarely provided. In this work we adapted the ''pellet culture'' method to obtain homogenous spheroids of MSC and maintain them in vitro for long term studies. We analysed their outer and inner structure over a 2-month period to provide morphological and architectural information regarding the spheroids generated. Quantitative and qualitative data were obtained using brightfield and confocal microscope imaging coupled to a computational analysis to estimate volume, sphericity, and jagging degree. In addition, histological evaluation was performed to more thoroughly assess the cellular composition and the internal architecture of the 3D spheroids. The results provided show that MSC spheroids generated with the proposed approach are homogeneous and stable, from both morphological and architectural points of view, for a period of at least 15 days, approximately between day 15 and day 30 after their generation. Accordingly, the approach proposed serves as a rapid, cost-effective, and efficient method to generate and maintain MSC spheroids using common entry-level laboratory equipment only.Keywords Mesenchymal stromal/stem cells Á 3D multicellular spheroids Á Pellet culture method Á Morphological analysis Á Microscopy Á Computational analysis Chiara Bellotti and Serena Duchi contributed equally to this work.

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Section: Msc Isolation and Culturementioning

confidence: 99%

Long term morphological characterization of mesenchymal stromal cells 3D spheroids built with a rapid method based on entry-level equipment

et al. 2016

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“…This allows comparison from various studies or of the yields from different tissues, but does not evaluate the capability of these cells to undergo other types of differentiation. Some investigations have reported the findings with respect to the number of phenotypic cfu when cultured under conditions whereby multi-lineage differentiation occurs (13). Information for selective differentiation of cultures is reported as Alizarin Red staining cfu for osteogenesis, Alcian Blue-positive staining cfu for chondrogenesis or Oil Red O staining cfu for adipogenesis (Figure 1).…”

Section: Harvesting and Characterization Of Msc From Different Tissuesmentioning

confidence: 99%

Stem Cell Considerations for the Clinician

Hasty

Cho

2016

Physical Medicine and Rehabilitation Clinics of North America

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Summary While there is ample evidence that beneficial results can be obtained from the use of MSC, several questions regarding their use remain to be answered. A better definition is needed for the mechanism/s by which the MSC are eliciting the positive outcome of their use whether it lies in the paracrine factors that are made by the MSC that affect the innate or adaptive immunity or impact the metabolism of resident cells, or their contribution as precursor cells giving rise to more differentiated reparative cells. If paracrine effects are desirable, commercial sources of allogeneic MSC could provide an easily obtainable, well-characterized reagent for the clinician. In their function as reparative cells, the longevity and fate of allogeneic MSC needs to be further evaluated on longer-term basis. The use of autologous cells has both advantages and its own inherent disadvantages involving additional procedures for collection, preparation and characterization of the MSC. It is apparent that a critical number of MSC are necessary for an optimal outcome for implantation. It remains to be determined if tissues such as adipose tissue provide more MSC than bone marrow and if the capability for differentiation is equivalent. A better understanding is also needed for how individual patient factors such as age, sex, drug regimens and co-morbidities influence the MSC population. For many of these questions, preclinical models will be helpful, but the task of evaluating and implementing these findings for orthopaedic patients falls onto the shoulders of clinical researchers. Evaluation of these questions is a daunting, but such a challenge fits the concept of personalized medicine in today’s medicine.

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“…In the past, these cells would have been expanded through culture in the lab, but the more recent impetus was to condense the harvest and implantation down to one surgical procedure. Currently, the standard is aspiration from the posterior iliac crest [15,16], as this site is potentially psychologically less traumatic [16] and is easily accessible with the needle. Compared to harvesting from the anterior pelvis, there is no difference in the total number of cells obtained, but there are more connective tissue progenitors from a posterior aspiration [15].…”

Section: Biologic Adjuvantsmentioning

confidence: 99%

“…Currently, the standard is aspiration from the posterior iliac crest [15,16], as this site is potentially psychologically less traumatic [16] and is easily accessible with the needle. Compared to harvesting from the anterior pelvis, there is no difference in the total number of cells obtained, but there are more connective tissue progenitors from a posterior aspiration [15]. After spinning the cells down, between 0.001 and 0.01 % of the cells obtained were found to be MSCs, with the total number of viable cells obtained decreasing with age [17].…”

Section: Biologic Adjuvantsmentioning

confidence: 99%

Biologic adjuvants and bone: current use in orthopedic surgery

Smith

Goldstein

Ekstein

2015

Curr Rev Musculoskelet Med

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Normal bone healing is a complex process that eventually restores original structure and function to the site of trauma. However, clinical circumstances such as nonunion, critical-sized defects, systemic bone disease, and fusion procedures have stimulated a search for ways to enhance this normal healing process. Biologics are an important part of this search and many, including bone marrow aspirate concentrate, demineralized bone matrix, platelet-rich plasma, bone morphogenic proteins, and platelet-derived growth factor, are currently in clinical use. Many others, including mesenchymal stem cells, parathyroid hormone, and Nel-like molecule-1 (NELL-1) will likely be in use in the future depending on the results of preclinical and clinical trials.

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The Posterior Iliac Crest Outperforms the Anterior Iliac Crest When Obtaining Mesenchymal Stem Cells from Bone Marrow

Abstract: The harvesting of bone marrow from the posterior iliac crest appears to be preferred, as it provided a modestly higher concentration of colony-founding connective-tissue progenitors than comparable aspirate from the anterior iliac crest.

Cited by 95 publications

References 3 publications

Long term morphological characterization of mesenchymal stromal cells 3D spheroids built with a rapid method based on entry-level equipment

Long term morphological characterization of mesenchymal stromal cells 3D spheroids built with a rapid method based on entry-level equipment

Stem Cell Considerations for the Clinician

Biologic adjuvants and bone: current use in orthopedic surgery

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