Culture Conditions for Primary Human Limbal Epithelial Cells

Osei-Bempong, Charles; Henein, Christin; Ahmad, Sajjad

doi:10.2217/rme.09.7

Cited by 23 publications

(14 citation statements)

References 51 publications

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“…Intriguingly, 3T3-J2 feeders have been used to culture diverse cells types, including epidermal keratinocytes (Rheinwald and Green, 1975a, Rheinwald and Green, 1975b), corneal epithelium (Osei-Bempong et al., 2009, Rama et al., 2010), and intestinal stem cells (Wang et al., 2015), among others. Nonetheless, the mechanism(s) by which 3T3-J2 feeders stabilize cultured progenitors are unknown.…”

Section: Discussionmentioning

confidence: 99%

Long-Term Culture of Self-renewing Pancreatic Progenitors Derived from Human Pluripotent Stem Cells

et al. 2017

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SummaryPluripotent stem cells have been proposed as an unlimited source of pancreatic β cells for studying and treating diabetes. However, the long, multi-step differentiation protocols used to generate functional β cells inevitably exhibit considerable variability, particularly when applied to pluripotent cells from diverse genetic backgrounds. We have developed culture conditions that support long-term self-renewal of human multipotent pancreatic progenitors, which are developmentally more proximal to the specialized cells of the adult pancreas. These cultured pancreatic progenitor (cPP) cells express key pancreatic transcription factors, including PDX1 and SOX9, and exhibit transcriptomes closely related to their in vivo counterparts. Upon exposure to differentiation cues, cPP cells give rise to pancreatic endocrine, acinar, and ductal lineages, indicating multilineage potency. Furthermore, cPP cells generate insulin+ β-like cells in vitro and in vivo, suggesting that they offer a convenient alternative to pluripotent cells as a source of adult cell types for modeling pancreatic development and diabetes.

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Section: Discussionmentioning

confidence: 99%

Long-Term Culture of Self-renewing Pancreatic Progenitors Derived from Human Pluripotent Stem Cells

et al. 2017

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“…Culture conditions for human limbal epithelial cells have recently been reviewed [Osei-Bempong et al, 2009]. The main differences between studies were whether the explant or cell suspension method was used, the usage or not of 3T3 mouse fibroblasts to co-culture, the type of substrate, and the culture medium used (Table I).…”

Section: Suspension Versus Explant Culturementioning

confidence: 99%

13 years of cultured limbal epithelial cell therapy: A review of the outcomes

Baylis

Figueiredo

Henein

et al. 2011

J of Cellular Biochemistry

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222

219

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The cornea is the clear tissue at the front of the eye which enables the transmission of light to the retina for normal vision. The surface of the cornea is composed of an epithelium which is renewed by stem cells located at the periphery of the cornea, a region known as the limbus. These limbal stem cells can become deficient as a result of various diseases of the eye's surface, resulting in the blinding disease of limbal stem cell deficiency. The treatment of this disease is often difficult and complex. In 1997, it was proposed that a small amount of limbal tissue containing limbal stem cells could be culture expanded and then transplanted. Since then various case reports and case series have been reported showing promising results. Here, we review the outcomes of this procedure over the past 13 years with the aim of highlighting the best culture and surgical techniques to date.

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“…Then, either the explant is used intact for culture or the limbal epithelial cells are digested from the explant to produce a cell suspension. The explant or suspension is then cultured for a period of time either on inactivated 3T3 mouse fibroblasts or on human amniotic membrane [31]. The overall result is the expansion of limbal epithelial cells, including the stem cells.…”

Section: Limbal Tissue Transplantation and Limbal Epithelial Cell Thementioning

confidence: 99%

Concise Review: Limbal Stem Cell Deficiency, Dysfunction, and Distress

Ahmad

2012

Stem Cells Translational Medicine

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129

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The cornea is the clear tissue at the front of the eye that transmits light to the retina at the back of the eye. The cornea is covered by an epithelium and surrounded by a narrow band of tissue known as the limbus. The limbus has two important roles in maintaining a healthy corneal epithelium. First, stem cells for the corneal epithelium reside at the limbus and not in the cornea. Second, the limbus acts as a barrier separating the clear avascular corneal epithelium from the surrounding vascular conjunctival tissue. A failure of these limbal functions can result in the painful and blinding disease of limbal stem cell deficiency. In this disease, the corneal epithelium cannot be maintained by the stem cells, and the corneal surface becomes replaced by hazy conjunctival tissue. There are many causes of limbal stem cell deficiency, such as burns to the eye, inflammatory diseases, and hereditary diseases. Current understanding of the pathophysiology of the disease is discussed here. In particular, understanding whether the limbal stem cells are lost or become dysfunctional or indeed whether the limbal microenvironment is disturbed is important when developing appropriate management strategies for the disease.

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Culture Conditions for Primary Human Limbal Epithelial Cells

Cited by 23 publications

References 51 publications

Long-Term Culture of Self-renewing Pancreatic Progenitors Derived from Human Pluripotent Stem Cells

Long-Term Culture of Self-renewing Pancreatic Progenitors Derived from Human Pluripotent Stem Cells

13 years of cultured limbal epithelial cell therapy: A review of the outcomes

Concise Review: Limbal Stem Cell Deficiency, Dysfunction, and Distress

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