Comparison of clinical grade human platelet lysates for cultivation of mesenchymal stromal cells from bone marrow and adipose tissue

Juhl, Morten; Tratwal, Josefine; Follin, Bjarke; Søndergaard, Rebekka Harary; Kirchhoff, Maria; Ekblond, Annette; Kastrup, Jens; Haack‐Sørensen, Mandana

doi:10.3109/00365513.2015.1099723

Cited by 45 publications

(43 citation statements)

References 37 publications

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“…Further, both Bieback et al (2009) andJuhl et al (2016) report that HPL supported long term monolayer expansion whilst still maintaining the MSCs phenotype and their differentiation potential. Given these positive data, herein it was established that Stemulate™ was better than FBS at increasing chondrocyte proliferation in monolayer culture.…”

Section: Discussionmentioning

confidence: 95%

“…There is clear evidence to confirm that 5-10 % (v/v) HPL is more effective in supporting monolayer expansion of human MSCs than both FBS and AS at the same concentration range (Bieback et al, 2009;Juhl et al, 2016;Trojahn Kølle et al, 2013). Further, both Bieback et al (2009) andJuhl et al (2016) report that HPL supported long term monolayer expansion whilst still maintaining the MSCs phenotype and their differentiation potential.…”

Section: Discussionmentioning

confidence: 98%

“…Given that Stemulate™ is used to expand a range of cell types, including adipose-derived mesenchymal stem cells (MSCs) and bone-marrow MSCs, it could be an alternative supplement to increase chondrocyte proliferation for cartilage cell therapy (Badowski et al, 2017;Juhl et al, 2016;Mangum et al, 2017;Riis et al, 2016;Søndergaard et al, 2017). Stemulate™ could reduce time in monolayer culture and reduce exposure to tissue-culture plastic, which in turn could reduce the risk of de-differentiation.…”

Section: Introductionmentioning

confidence: 99%

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Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

Sykes¹,

Kuiper²,

Richardson³

et al. 2018

eCM

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High hopes have been pinned on regenerative medicine strategies in order to prevent the progression of cartilage damage to osteoarthritis, particularly by autologous chondrocyte implantation (ACI). The loss of chondrocyte phenotype during in vitro monolayer expansion, a necessary step to obtain sufficient cell numbers, may be a key limitation in ACI. In this study, it was determined whether a shorter monolayer expansion approach could improve chondrogenic differentiation. The effects of two supplement types, foetal bovine serum (FBS) and Stemulate™ (a commercial source of human platelet lysate), on the expansion and re-differentiation potential of human chondrocytes, isolated from five individuals, were compared. Chondrocytes were expanded with 10 % FBS or 10 % Stemulate™. Pellets were cultured for 28 d in chondrogenic differentiation medium and assessed for the presence of cartilage matrix molecules and genes associated with chondrogenicity. Stemulate™ significantly enhanced the proliferation rate [average population doubling times: FBS, 25.07 ± 6.98 d (standard error of the mean, SEM) vs. Stemulate™, 13.10 ± 2.57 d (SEM)]. Sulphated glycosaminoglycans (sGAG), total collagen and qRT-PCR analyses of cartilage genes showed that FBS-expanded chondrocytes demonstrated significantly better chondrogenic capacity than Stemulate™-expanded chondrocytes. Histologically, FBS-expanded chondrocyte pellets appeared to be more stable, with a more intense staining for toluidine blue, indicating a greater chondrogenic capacity. Although Stemulate™ positively influenced chondrocyte proliferation, it had a negative effect on chondrogenic differentiation potential. This suggested that, in the treatment of cartilage defects, Stemulate™ might not be the ideal supplement for expanding chondrocytes (which maintained a chondrocyte phenotype) and, hence, for cell therapies (including ACI).

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

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Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

Sykes¹,

Kuiper²,

Richardson³

et al. 2018

eCM

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“…Surprisingly, no observable degree of adipogenic differentiation occurred in either of the cell populations. In our hands, human MSCs readily display lipid accumulation after short periods, 40 while this is difficult to attain with porcine MSCs even with prolonged induction periods (personal observation). Whether this is related to species variation or a unique attribute of SEER cells remains uncertain.…”

Section: Discussionmentioning

confidence: 84%

Identification of SSEA-1 expressing enhanced reprogramming (SEER) cells in porcine embryonic fibroblasts

et al. 2017

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Previous research has shown that a subpopulation of cells within cultured human dermal fibroblasts, termed multilineage-differentiating stress enduring (Muse) cells, are preferentially reprogrammed into induced pluripotent stem cells. However, controversy exists over whether these cells are the only cells capable of being reprogrammed from a heterogeneous population of fibroblasts. Similarly, there is little research to suggest such cells may exist in embryonic tissues or other species. To address if such a cell population exists in pigs, we investigated porcine embryonic fibroblast populations (pEFs) and identified heterogeneous expression of several key cell surface markers. Strikingly, we discovered a small population of stage-specific embryonic antigen 1 positive cells (SSEA-1C) in Danish Landrace and G€ ottingen minipig pEFs, which were absent in the Yucatan pEFs. Furthermore, reprogramming of SSEA-1C sorted pEFs led to higher reprogramming efficiency. Subsequent transcriptome profiling of the SSEA-1C vs. the SSEA-1neg cell fraction revealed highly comparable gene signatures. However several genes that were found to be upregulated in the SSEA-1C cells were similarly expressed in mesenchymal stem cells (MSCs). We therefore termed these cells SSEA-1 Expressing Enhanced Reprogramming (SEER) cells. Interestingly, SEER cells were more effective at differentiating into osteocytes and chondrocytes in vitro. We conclude that SEER cells are more amenable for reprogramming and that the expression of mesenchymal stem cell genes is advantageous in the reprogramming process. This data provides evidence supporting the elite theory and helps to delineate which cell types and specific genes are important for reprogramming in the pig.

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“…Adipose derived stromal cells P2 harvested from a Quantum system and flasks were tested for their ability to differentiate into osteoblasts, adipocytes and chondrocytes lineages, as described in our previous publication using StemPro differentiation kit (Gibco, Life Technology), according to the manufacturer’s protocols [18]. …”

Section: Methodsmentioning

confidence: 99%

Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture

et al. 2016

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BackgroundAdipose derived stromal cells (ASCs) are a rich and convenient source of cells for clinical regenerative therapeutic approaches. However, applications of ASCs often require cell expansion to reach the needed dose. In this study, cultivation of ASCs from stromal vascular fraction (SVF) over two passages in the automated and functionally closed Quantum Cell Expansion System (Quantum system) is compared with traditional manual cultivation.MethodsStromal vascular fraction was isolated from abdominal fat, suspended in α-MEM supplemented with 10% Fetal Bovine Serum and seeded into either T75 flasks or a Quantum system that had been coated with cryoprecipitate. The cultivation of ASCs from SVF was performed in 3 ways: flask to flask; flask to Quantum system; and Quantum system to Quantum system. In all cases, quality controls were conducted for sterility, mycoplasmas, and endotoxins, in addition to the assessment of cell counts, viability, immunophenotype, and differentiation potential.ResultsThe viability of ASCs passage 0 (P0) and P1 was above 96%, regardless of cultivation in flasks or Quantum system. Expression of surface markers and differentiation potential was consistent with ISCT/IFATS standards for the ASC phenotype. Sterility, mycoplasma, and endotoxin tests were consistently negative. An average of 8.0 × 107 SVF cells loaded into a Quantum system yielded 8.96 × 107 ASCs P0, while 4.5 × 106 SVF cells seeded per T75 flask yielded an average of 2.37 × 106 ASCs—less than the number of SVF cells seeded. ASCs P1 expanded in the Quantum system demonstrated a population doubling (PD) around 2.2 regardless of whether P0 was previously cultured in flasks or Quantum, while ASCs P1 in flasks only reached a PD of 1.0. Conclusion: Manufacturing of ASCs in a Quantum system enhances ASC expansion rate and yield significantly relative to manual processing in T-flasks, while maintaining the purity and quality essential to safe and robust cell production. Notably, the use of the Quantum system entails significantly reduced working hours and thereby costs.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-016-1080-9) contains supplementary material, which is available to authorized users.

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Comparison of clinical grade human platelet lysates for cultivation of mesenchymal stromal cells from bone marrow and adipose tissue

Abstract: All three clinically approved commercial human platelet lysates accelerated proliferation of BMSCs and ASCs and the cells meet the ISCT mesenchymal phenotypic requirements without exhibiting chromosomal aberrations.

Cited by 45 publications

References 37 publications

Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

Impact of human platelet lysate on the expansion and chondrogenic capacity of cultured human chondrocytes for cartilage cell therapy

Identification of SSEA-1 expressing enhanced reprogramming (SEER) cells in porcine embryonic fibroblasts

Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture

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